Feline immunoglobulin e molecules and related methods

ABSTRACT

The invention relates to: nucleic acid molecules encoding the light chain and heavy chain of feline immunoglobulin E (IgE), including species-specific regions of feline IgE; proteins encoded by the nucleic acid molecules; inhibitors to the nucleic acids and proteins; antibodies to the proteins; cells transformed with the nucleic acid molecules; assays employing the transformed cells, nucleic acids, antibodies and/or proteins or portions thereof; methods for treating IgE-mediated responses (ie. allergy) using the materials provided; methods for eliciting an immune response to IgE and kits containing the nucleic acid molecules, proteins or derivatives thereof (ie. antibodies).

FIELD OF THE INVENTION

[0001] The present invention relates to the field of feline IgE-mediatedresponses, and materials and methods useful to alter natural processrelated to IgE-mediated responses. The present invention thereforerelates to vaccine technology, small molecule/antibody technology,molecular biology tools, and immunological techniques related to felineIgE and its function.

BACKGROUND OF THE INVENTION

[0002] Allergic responses in mammals are known to be mediated byimmunoglobulin E. IgE molecules bind the Fcε receptor on mast cells and,when complexed with antigen, trigger a cascade of events that leads tothe release of allergic mediators (ie. histamine, prostaglandins andproteases). Thus, interference with the IgE/Fcε receptor interaction isan avenue for controlling allergic responses. Interference with the IgEantibody/Fcε receptor interaction will also affect the pathology ofatopic disease, hyper IgE syndrome, internal parasite infections and Bcell neoplasia.

[0003] The species-specific portion of the IgE, the IgE constant region(on the heavy chain and involved in Fcε receptor binding) is ofparticular importance in design and manufacture of compounds useful tointerfere with the IgE/Fcε receptor interaction, because compounds whichare specific for this region produce little interference withnon-IgE/receptor interactions. Moreover, the IgE constant region can beutilized in the design and manufacture of vaccines useful to elicitspecies- and immunoglobulin-specific anti-IgE immune responses.

[0004] The DNA and amino acid sequences of IgE molecules from severalspecies, including human, rat, mouse and dog, have been reported.Peptides derived from known IgE sequences have been used to generateantibodies which alter IgE function. U.S. Pat. No. 5,091,313 is directedto the prevention or control of IgE-mediated allergic symptoms throughthe use of monoclonal or polyclonal antibodies raised against epitopespresent in B cell-associated or soluble human IgE. WO90/15878 disclosesthe use of peptides derived from human, rat or mouse IgE sequences togenerate antibodies which inhibit IgE-mediated mast cell degranulation.U.S. Pat. No. 4,223,016 discloses the use of peptides derived from IgEsequences for allergic desensitization. U.S. Pat. No. 5,629,415discloses the canine IgE sequence and uses therefor.

SUMMARY OF THE INVENTION

[0005] The present invention provides isolated nucleic acid moleculeswhich encode a portion of the heavy chain of feline IgE, isolatedproteins encoded by the nucleic acid molecules, recombinant constructsand cells comprising the nucleic acid compounds and/or proteins,antibodies to the isolated proteins, therapeutic compositions useful fortreating feline IgE-mediated responses (including i.e., vaccines),methods for treating feline IgE-mediated responses, methods foreliciting a feline IgE-mediated immune response, and kits comprising thematerials provided. The present invention also provides nucleic acidmolecules, proteins and methods related to the feline IgE light chain.

[0006] The present invention therefore provides isolated nucleic acidmolecules encoding a portion of a feline IgE heavy chain molecule,wherein said nucleic acid molecules comprise a nucleic acid sequenceselected from the group consisting of:

[0007] (a) a nucleic acid sequence which has more than 82% identity to anucleic acid sequence selected from the group consisting of: SEQ ID NO1; and SEQ ID NO 28, wherein said identity can be determined using theDNAsis computer program and default parameters;

[0008] (b) a nucleic acid sequence which encodes a feline heavy chainprotein which has more than 76% identity to an amino acid sequenceselected from the group consisting of: SEQ ID NO 2; and SEQ ID NO 29,wherein said identity can be determined using the DNAsis computerprogram and default parameters;

[0009] (c) a nucleic acid sequence which encodes and a feline heavychain protein encoded by an allelic variant of a nucleic acid sequenceselected from the group consisting of: SEQ ID NO 1; and SEQ ID NO 28;and

[0010] (d) a nucleic acid sequence which has more than 90% identity to anucleic acid sequence selected from the group consisting of: SEQ ID NO3; SEQ ID NO 4; SEQ ID NO 6; SEQ ID NO 7; SEQ ID NO 9; SEQ ID NO 10; SEQID NO 12; SEQ ID NO 13; SEQ ID NO 15; SEQ ID NO 16; SEQ ID NO 18; andSEQ ID NO 31; and

[0011] (e) a nucleic acid molecule fully complementary to a nucleic acidmolecule selected from the group consisting of: a nucleic acid moleculeof (a); a nucleic acid molecule of (b); and a nucleic acid molecule of(c).

[0012] The preferred nucleic acid molecules are those with immunologicalsignificance. At the time of filing, nucleic acid molecules which encodethose which encode the constant region, specifically those which encodea Fcε receptor (sometimes called “FcεR”) binding region, is preferred.In particular, nucleic acid molecules which encode a feline IgE Fcεreceptor binding region and which comprises SEQ ID NO 4, SEQ ID NO 7 orSEQ ID NO 10 are preferred. Also provided is a nucleic acid moleculewhich encodes a feline IgE constant region and comprises SEQ ID NO 13.

[0013] The present invention also provides nucleic acid molecules whichencode a feline IgE light chain protein and which comprise a nucleicacid molecule which encodes a protein with more than 84% identity to SEQID NO 19, with a nucleic acid molecule which comprises SEQ ID NO 19being preferred.

[0014] The present invention also comprises expression vectors andrecombinant cells comprising the present nucleic acid molecules. Alsoprovided are fusion protein constructs comprising the present nucleicacid compounds.

[0015] The present invention also comprises isolated proteins encoding aportion of a feline IgE heavy chain molecule, wherein said proteinscomprise an amino acid sequence selected from the group consisting of:

[0016] (a) an amino acid sequence encoded by a nucleic acid sequencewhich has more than 82% identity to a nucleic acid sequence selectedfrom the group consisting of: SEQ ID NO 1; and SEQ ID NO: 28, whereinsaid identity can be determined using the DNAsis computer program anddefault parameters;

[0017] (b) an amino acid sequence which has more than 76% identity to anamino acid sequence selected from the group consisting of: SEQ ID NO 2;and SEQ ID NO: 29, wherein said identity can be determined using theDNAsis computer program and default parameters;

[0018] (c) an amino acid sequence encoded by an allelic variant of anucleic acid sequence selected from the group consisting of: SEQ ID NO1; and SEQ ID NO: 28; and

[0019] (d) an amino acid sequence encoded by a a nucleic acid sequencewhich has more than 90% identity to a nucleic acid sequence selectedfrom the group consisting of: SEQ ID NO 3; SEQ ID NO 4; SEQ ID NO 6; SEQID NO 7; SEQ ID NO 9; SEQ ID NO 10; SEQ ID NO 12; SEQ ID NO 13; SEQ IDNO 15; SEQ ID NO 16; SEQ ID NO 18; SEQ ID NO 28; and SEQ ID NO 30.

[0020] The preferred embodiments of this aspect of the present inventioninclude those proteins capable of binding to Fc receptor, in particular,SEQ ID NO 5, SEQ ID NO 8, SEQ ID NO 11 and SEQ ID NO 14.

[0021] In another embodiment, there are provided antibodies selectivefor a protein of the present invention. In particular, antibodiesdesignated H-100, H-101, H-102, H-103, H-106 are preferred.

[0022] In another embodiment, there are provided therapeuticcompositions useful for inhibiting an immune response to feline IgE,wherein said therapeutic composition is selected from the groupconsisting of:

[0023] (a) a nucleic acid molecule of the present invention;

[0024] (b) a protein encoded by a nucleic acid of (a);

[0025] (c) an inhibitor of a nucleic acid of (a); and

[0026] (d) an inhibitor of a protein of (b).

[0027] Preferred embodiments of this aspect of the present invention areantibodies selective for the proteins of the present invention, inparticular, H-100, H-101, H-102, H-103, H-106 are preferred.

[0028] Also provided by the present invention are methods to identifythe ability of a test compound to interfere with IgE/Fcε, interaction,comprising: contacting the test compound with a protein of the presentinvention; and determining whether the test compound and said proteininteract.

[0029] Also provided by the present invention are methods for inhibitingan immune response to feline IgE, comprising administering at least onetherapeutic composition of the present invention.

[0030] Also provided by the present invention are diagnostic kits,comprising a container comprising a at least one composition selectedfrom the group consisting of:

[0031] (a) a nucleic acid molecule of the present invention;

[0032] (b) a protein encoded by a nucleic acid of (a);

[0033] (c) an inhibitor of a nucleic acid of (a); and

[0034] (d) an inhibitor of a protein of (b).

[0035] Also provided are isolated nucleic acid molecules encoding aportion of a feline IgE light chain protein, wherein said nucleic acidmolecule comprises a nucleic acid sequence selected from the groupconsisting of:

[0036] (a) a nucleic acid sequence which has more than 84% identity toSEQ ID NO 19, and wherein said identity can be determined using theDNAsis computer program and default parameters;

[0037] (b) a nucleic acid sequence which encodes a feline heavy chainprotein selected from the group consisting of: a feline heavy chainprotein which has more than 61% identity to SEQ ID NO 20, wherein saididentity can be determined using the DNAsis computer program and defaultparameters; and a feline heavy chain protein encoded by an allelicvariant of SEQ ID NO 19;

[0038] (c) a nucleic acid sequence which has more than 95% identity to anucleic acid sequence selected from the group consisting of: SEQ ID NO23; and SEQ ID NO 25; and

[0039] (d) a nucleic acid molecule fully complementary to a nucleic acidmolecule selected from the group consisting of: a nucleic acid moleculeof (a); a nucleic acid molecule of (b); and a nucleic acid molecule of(c).

[0040] Also provided are isolated proteins encoding a portion of afeline IgE light chain molecule, wherein said protein comprises an aminoacid sequence selected from the group consisting of:

[0041] (a) an amino acid sequence encoded by a nucleic acid sequencewhich has more than 84% identity to SEQ ID NO 19, and wherein saididentity can be determined using the DNAsis computer program and defaultparameters;

[0042] (b) an amino acid sequence which has more than 61% identity toSEQ ID NO 20, wherein said identity can be determined using the DNAsiscomputer program and default parameters;

[0043] (c) an amino acid sequence encoded by an allelic variant of SEQID NO 19; and

[0044] (d) a nucleic acid sequence which has more than 95% identity to anucleic acid sequence selected from the group consisting of: SEQ ID NO23; and SEQ ID NO 25.

[0045] Definitions

[0046] “Allelic variant” is meant to refer to a full length gene orpartial sequence of a full length gene that occurs at essentially thesame locus (or loci) as the referent sequence, but which, due to naturalvariations caused by, for example, mutation or recombination, has asimilar but not identical sequence. Allelic variants typically encodeproteins having similar activity to that of the protein encoded by thegene to which they are being compared. Allelic variants can alsocomprise alterations in the 5′ or 3′ untranslated regions of the gene(e.g., in regulatory control regions).

[0047] “Antibody” as used herein includes both polyclonal and monoclonalantibodies as well as fragments thereof, such as Fv, Fab and F(ab)₂fragments that are capable of binding antigen or hapten.

[0048] “Fcε receptor” means any Fcε receptor recognized in the art,including the “low” affinity or “high” affinity receptors, or any suchnew receptors discovered.

[0049] “Feline Fcε receptor binding region” means a region of the felineIgE molecule that is capable of binding to a Fcε receptor, including theentire, naturally-occurring binding region, portions thereof that bindto the Fcε receptor, or modifications of either the entirenaturally-occurring binding region or portions thereof.

[0050] “Feline IgE-mediated immune response” means not only any humoralor cellular immune response, but also any biological response resultingfrom an IgE/Fcε receptor interaction.

[0051] “Fragment” is meant to refer to any subset of the referentnucleic acid molecule.

[0052] “Incite” means causing any affect, ie. stimulation, of the felineIgE-mediated immune response.

[0053] “Immunogenic amounts” means at least the minimal amount necessaryto incite a feline IgE-mediated immune response.

[0054] “Proteins” means any compounds which comprise amino acids,including peptides, polypeptides, fusion proteins, etc.

[0055] Moreover, for the purposes of the present invention, the term “a”or “an” entity refers to one or more of that entity; for example, “aprotein” or “a nucleic acid molecule” refers to one or more of thosecompounds or at least one compound. As such, the terms “a” (or “an”),“one or more” and “at least one” can be used interchangeably herein. Itis also to be noted that the terms “comprising”, “including”, and“having” can be used interchangeably. Furthermore, a compound “selectedfrom the group consisting of” refers to one or more of the compounds inthe list that follows, including mixtures (i.e., combinations) of two ormore of the compounds. According to the present invention, an isolated,or biologically pure, protein or nucleic acid molecule is a compoundthat has been removed from its natural milieu. As such, “isolated” and“biologically pure” do not necessarily reflect the extent to which thecompound has been purified. An isolated compound of the presentinvention can be obtained from its natural source, can be produced usingmolecular biology techniques or can be produced by chemical synthesis.

DETAILED DESCRIPTION OF THE INVENTION

[0056] The present invention provides isolated nucleic acid moleculeswhich encode a portion of the heavy chain of feline IgE, isolatedproteins encoded by the nucleic acid molecules, recombinant constructsand cells comprising the nucleic acid compounds and/or proteins,antibodies to the isolated proteins, inhibitors of the proteins andnucleic acids, therapeutic compositions useful for treating felineIgE-mediated responses (including i.e., vaccines), methods for treatingfeline IgE-mediated responses, methods for eliciting a felineIgE-mediated immune response, and kits comprising the materialsprovided. The present invention also provides feline IgE light chainnucleic- and amino acid molecules, and associated materials.

[0057] The present invention therefore provides isolated nucleic acidmolecules encoding a portion of a feline heavy chain molecule, whereinsaid nucleic acid molecules have more than 82% identity to SEQ ID NO 1and/or SEQ ID NO 28, and wherein said identity can be determined usingthe DNAsis computer program and default parameters, as well as nucleicacid molecules fully complementary to those nucleic acid molecules.

[0058] Moreover, there is provided isolated nucleic acid moleculesencoding a portion of a feline IgE heavy chain molecule, wherein saidnucleic acid molecule comprises a nucleic acid sequence selected fromthe group consisting of:

[0059] (a) a nucleic acid sequence which has more than 82% identity to anucleic acid sequence selected from the group consisting of: SEQ ID NO1; and SEQ ID NO 28, wherein said identity can be determined using theDNAsis computer program and default parameters;

[0060] (b) a nucleic acid sequence which encodes a feline heavy chainprotein which has more than 76% identity to an amino acid sequenceselected from the group consisting of: SEQ ID NO 2; and SEQ ID NO 29,wherein said identity can be determined using the DNAsis computerprogram and default parameters;

[0061] (c) a nucleic acid sequence which encodes and a feline heavychain protein encoded by an allelic variant of a nucleic acid sequenceselected from the group consisting of: SEQ ID NO 1; and SEQ ID NO 28;and

[0062] (d) a nucleic acid sequence which has more than 90% identity to anucleic acid sequence selected from the group consisting of: SEQ ID NO3; SEQ ID NO 4; SEQ ID NO 6; SEQ ID NO 7; SEQ ID NO 9; SEQ ID NO 10; SEQID NO 12; SEQ ID NO 13; SEQ ID NO 15; SEQ ID NO 16; SEQ ID NO 18; andSEQ ID NO 31; and

[0063] (e) a nucleic acid molecule fully complementary to a nucleic acidmolecule selected from the group consisting of: a nucleic acid moleculeof (a); a nucleic acid molecule of (b); and a nucleic acid molecule of(c).

[0064] Allelic variants, fragments and homologues are, by definition of“nucleic acid molecule”, included within this and other embodiments.

[0065] The preferred nucleic acid molecules are those with immunologicsignificance. At the time of filing, nucleic acids which encode theconstant region, specifically those nucleic acid molecules which encodethe Fcε receptor binding region, are preferred. In particular, SEQ ID NO4, SEQ ID NO 7, SEQ ID NO 10, and SEQ ID NO 13 and complements thereofare most preferred.

[0066] The present invention also provides nucleic acid molecules whichencode a feline IgE light chain protein and which comprise a nucleicacid molecule which encodes a protein with more than 84% identity to SEQID NO 19, with a nucleic acid molecule which comprises SEQ ID NO 19being preferred.

[0067] The present invention also comprises expression vectors andrecombinant cells comprising the present nucleic acid molecules. Alsoprovided are fusion proteins constructed using the present nucleic acidcompounds.

[0068] Included within the scope of the present invention, withparticular regard to the nucleic acids above, are allelic variants,degenerate sequences and homologues. Allelic variants are well known tothose skilled in the art and would be expected to be found within agiven cat since the genome is diploid and/or among a group of two ormore cats. The present invention also includes variants due tolaboratory manipulation, such as, but not limited to, variants producedduring polymerase chain reaction amplification or site directedmutagenesis. It is also well known that there is a substantial amount ofredundancy in the various codons which code for specific amino acids.Therefore, this invention is also directed to those nucleic acidsequences which contain alternative codons which code for the eventualtranslation of the identical amino acid. Also included within the scopeof this invention are mutations either in the nucleic acid sequence orthe translated protein which do not substantially alter the ultimatephysical properties of the expressed protein. For example, substitutionof valine for leucine, arginine for lysine, or asparagine for glutaminemay not cause a change in functionality of the polypeptide. Lastly, anucleic acid sequence homologous to the exemplified nucleic acidmolecules (or allelic variants or degenerates thereof) will have atleast 85%, preferably 90%, and most preferably 95% sequence identitywith a nucleic acid molecule in the sequence listing.

[0069] Stringent hybridization conditions are determined based ondefined physical properties of the gene to which the nucleic acidmolecule is being hybridized, and can be defined mathematically.Stringent hybridization conditions are those experimental parametersthat allow an individual skilled in the art to identify significantsimilarities between heterologous nucleic acid molecules. Theseconditions are well known to those skilled in the art. See, for example,Sambrook, et al., 1989, Molecular Cloning: A Laboratory Manual, ColdSpring Harbor Labs Press, and Meinkoth, et al., 1984, Anal. Biochem.138, 267-284, each of which is incorporated by reference herein in itsentirety. As explained in detail in the cited references, thedetermination of hybridization conditions involves the manipulation of aset of variables including the ionic strength (M, in moles/liter), thehybridization temperature (° C.), the concentration of nucleic acidhelix destabilizing agents (such as formamide), the average length ofthe shortest hybrid duplex (n), and the percent G+C composition of thefragment to which an unknown nucleic acid molecule is being hybridized.For nucleic acid molecules of at least about 150 nucleotides, thesevariables are inserted into a standard mathematical formula to calculatethe melting temperature, or T_(m), of a given nucleic acid molecule. Asdefined in the formula below, T_(m) is the temperature at which twocomplementary nucleic acid molecule strands will disassociate, assuming100% complementarity between the two strands: T_(m)=81.5° C.+16.6 logM+0.41(%G+C)-500/n-0.61(% formamide). For nucleic acid molecules smallerthan about 50 nucleotides, hybrid stability is defined by thedissociation temperature (T_(d)), which is defined as the temperature atwhich 50% of the duplexes dissociate. For these smaller molecules, thestability at a standard ionic strength is defined by the followingequation: T_(d)=4(G+C)+2(A+T). A temperature of 5° C. below T_(d) isused to detect hybridization between perfectly matched molecules.

[0070] Also well known to those skilled in the art is how base-pairmismatch, i.e. differences between two nucleic acid molecules beingcompared, including non-complementarity of bases at a given location,and gaps due to insertion or deletion of one or more bases at a givenlocation on either of the nucleic acid molecules being compared, willaffect T_(m) or T_(d) for nucleic acid molecules of different sizes. Forexample, T_(m) decreases about 1° C. for each 1% of mismatchedbase-pairs for hybrids greater than about 150 bp, and T_(d) decreasesabout 5° C. for each mismatched base-pair for hybrids below about 50 bp.Conditions for hybrids between about 50 and about 150 base-pairs can bedetermined empirically and without undue experimentation using standardlaboratory procedures well known to those skilled in the art. Thesesimple procedures allow one skilled in the art to set the hybridizationconditions (by altering, for example, the salt concentration, theformamide concentration or the temperature) so that only nucleic acidhybrids with less than a specified % base-pair mismatch will hybridize.Stringent hybridization conditions are commonly understood by thoseskilled in the art to be those experimental conditions that will allowhybridization between molecules having about 30% or less base-pairmismatch (i.e., about 70% or greater identity). Because one skilled inthe art can easily determine whether a given nucleic acid molecule to betested is less than or greater than about 50 nucleotides, and cantherefore choose the appropriate formula for determining hybridizationconditions, he or she can determine whether the nucleic acid moleculewill hybridize with a given gene under stringent hybridizationconditions and similarly whether the nucleic acid molecule willhybridized under conditions designed to allow a desired amount of basepair mismatch.

[0071] Hybridization reactions are often carried out by attaching thenucleic acid molecule to be hybridized to a solid support such as amembrane, and then hybridizing with a labeled nucleic acid molecule,typically referred to as a probe, suspended in a hybridization solution.Examples of common hybridization reaction techniques include, but arenot limited to, the well-known Southern and northern blottingprocedures. Typically, the actual hybridization reaction is done undernon-stringent conditions, i.e., at a lower temperature and/or a highersalt concentration, and then high stringency is achieved by washing themembrane in a solution with a higher temperature and/or lower saltconcentration in order to achieve the desired stringency.

[0072] For example, if the skilled artisan wished to identify a nucleicacid molecule that hybridized under stringent hybridization conditionswith a feline nucleic acid molecule of about 150 bp in length, thefollowing conditions could preferably be used. The average G+C contentof feline genome is about 53%. The unknown nucleic acid molecules wouldbe attached to a support membrane, and the 150 bp probe would belabeled, e.g. with a radioactive tag. The hybridization reaction couldbe carried out in a solution comprising 2×SSC and 0% formamide, at atemperature of about 37° C. (low stringency conditions). Solutions ofdiffering concentrations of SSC can be made by one of skill in the artby diluting a stock solution of 20×SSC (175.3 gram NaCl and about 88.2gram sodium citrate in 1 liter of water, pH 7) to obtain the desiredconcentration of SSC. In order to achieve high stringency hybridization,the skilled artisan would calculate the washing conditions required toallow up to 30% base-pair mismatch. For example, in a wash solutioncomprising 1×SSC and 0% formamide, the T_(m) of perfect hybrids would beabout 86° C.:

81.5° C.+16.6 log (0.15M)+(0.53×39)−(500/150)−(0.61×0)=86.3° C.

[0073] Thus, to achieve hybridization with nucleic acid molecules havingabout 30% base-pair mismatch, hybridization washes would be carried outat a temperature of about 56° C. It is thus within the skill of one inthe art to calculate additional hybridization temperatures based on thedesired percentage base-pair mismatch, formulae and G/C contentdisclosed herein. For example, it is appreciated by one skilled in theart that as the nucleic acid molecule to be tested for hybridizationagainst nucleic acid molecules of the present invention having sequencesspecified herein becomes longer than 150 nucleotides, the T_(m) for ahybridization reaction allowing up to 30% base-pair mismatch will notvary significantly from 56° C.

[0074] It is known in the art that there are commercially availablecomputer programs for determining the degree of similarity between twonucleic acid sequences. These computer programs include various knownmethods to determine the percentage identity and the number and lengthof gaps between hybrid nucleic acid molecules. Preferred methods todetermine the percent identity among amino acid sequences and also amongnucleic acid sequences include analysis using one or more of thecommercially available computer programs designed to compare and analyzenucleic acid or amino acid sequences. These computer programs include,but are not limited to, GCG™ (available from Genetics Computer Group,Madison, Wis.), DNAsis™ (available from Hitachi Software, San Bruno,Calif.) and MacVector™ (available from the Eastman Kodak Company, NewHaven, Conn.). A preferred method to determine percent identity amongamino acid sequences and also among nucleic acid sequences includesusing the Compare function by maximum matching within the program DNAsisVersion 2.1 using default parameters.

[0075] In one embodiment of the present invention, a preferred felineIgE nucleic acid molecule includes an isolated nucleic acid moleculewhich is at least about 50 nucleotides, or at least about 150nucleotides, and which hybridizes under conditions which preferablyallow about 50% base pair mismatch, more preferably under conditionswhich allow about 45% base pair mismatch, more preferably underconditions which allow about 40% base pair mismatch, more preferablyunder conditions which allow about 35% base pair mismatch, morepreferably under conditions which allow about 30% base pair mismatch,more preferably under conditions which allow about 25% base pairmismatch, more preferably under conditions which allow about 20% basepair mismatch, more preferably under conditions which allow about 15%base pair mismatch, more preferably under conditions which allow about10% base pair mismatch and even more preferably under conditions whichallow about 5% base pair mismatch with a nucleic acid molecule selectedfrom the group consisting of SEQ ID NO 1, SEQ ID NO 4, SEQ ID NO 7, SEQID NO 10, SEQ ID NO 13, SEQ ID NO 16, and/or SEQ ID NO 19.

[0076] Another embodiment of the present invention includes a nucleicacid molecule comprising at least about 150 base-pairs, wherein thenucleic acid molecule hybridizes, in a solution comprising 1×SSC and 0%formamide, at a temperature of about 56° C., to a nucleic acid sequenceselected from the group consisting of: SEQ ID NO 1; SEQ ID NO 3; SEQ IDNO 4; SEQ ID NO 6; SEQ ID NO 7; SEQ ID NO 9; SEQ ID NO 10; SEQ ID NO 12;SEQ ID NO 13; SEQ ID NO 15; SEQ ID NO 16; and SEQ ID NO 18. Alsopreferred are fragments of any of such nucleic acid molecules.

[0077] Comparison of nucleic acid sequence SEQ ID NO 1 (i.e., thenucleic acid sequence of a portion of the feline IgE heavy chain) withnucleic acid sequences reported in GenBank indicates that SEQ ID NO 1showed the most homology, i.e. about 82% identity, between SEQ ID NO 1and a Canis familiaris IgE heavy chain region (Accession Number L36872).

[0078] Additional preferred feline IgE nucleic acid molecules of thepresent invention include an isolated nucleic acid molecule which is atleast about 50 nucleotides, or at least about 150 nucleotides,comprising a nucleic acid sequence that is preferably at least about 45%identical, more preferably about 50% identical, more preferably about55% identical, more preferably about 60% identical, more preferablyabout 65% identical, more preferably about 70% identical, morepreferably about 75% identical, more preferably about 80% identical,more preferably about 85% identical, more preferably about 90% identicaland even more preferably about 95% identical to a nucleic acid sequenceselected from the group consisting of SEQ ID NO 1, SEQ ID NO 4, SEQ IDNO 7, SEQ ID NO 10, SEQ ID NO 13, SEQ ID NO 16, and/or SEQ ID NO 19.Also preferred are fragments of any of such nucleic acid molecules.Percent identity may be determined using the Compare function by maximummatching within the program DNAsis Version 2.1 using default parameters.

[0079] Knowing the nucleic acid sequences of certain feline IgE nucleicacid molecules of the present invention allows one skilled in the artto, for example, (a) make copies of those nucleic acid molecules, (b)obtain nucleic acid molecules including at least a portion of suchnucleic acid molecules (e.g., nucleic acid molecules includingfull-length genes, full-length coding regions, regulatory controlsequences, truncated coding regions), and (c) obtain feline IgE nucleicacid molecules from other species. Such nucleic acid molecules can beobtained in a variety of ways including screening appropriate expressionlibraries with antibodies of the present invention; traditional cloningtechniques using oligonucleotide probes of the present invention toscreen appropriate libraries of DNA; and PCR amplification ofappropriate libraries or DNA using oligonucleotide primers of thepresent invention. Preferred libraries to screen or from which toamplify nucleic acid molecules include canine cDNA libraries as well asgenomic DNA libraries. Similarly, preferred DNA sources to screen orfrom which to amplify nucleic acid molecules include adult cDNA andgenomic DNA. Techniques to clone and amplify genes are disclosed, forexample, in Sambrook et al., ibid.

[0080] The present invention also includes nucleic acid molecules thatare oligonucleotides capable of hybridizing, under stringenthybridization conditions, with complementary regions of other,preferably longer, nucleic acid molecules of the present invention suchas those comprising feline IgE genes or other feline IgE nucleic acidmolecules. Oligonucleotides of the present invention can be RNA, DNA, orderivatives of either. The minimum size of such oligonucleotides is thesize required for formation of a stable hybrid between anoligonucleotide and a complementary sequence on a nucleic acid moleculeof the present invention. Minimal size characteristics are disclosedherein. The present invention includes oligonucleotides that can be usedas, for example, probes to identify nucleic acid molecules, primers toproduce nucleic acid molecules or therapeutic reagents to inhibit felineFcεRα protein production or activity (e.g., as antisense-, triplexformation-, ribozyme- and/or RNA drug-based reagents). The presentinvention also includes the use of such oligonucleotides to protectanimals from disease using one or more of such technologies. Appropriateoligonucleotide-containing therapeutic compositions can be administeredto an animal using techniques known to those skilled in the art.

[0081] Recombinant molecules of the present invention may also (a)contain secretory signals (i.e., signal segment nucleic acid sequences)to enable an expressed feline IgE protein of the present invention to besecreted from the cell that produces the protein and/or (b) containfusion sequences which lead to the expression of nucleic acid moleculesof the present invention as fusion proteins. Examples of suitable signalsegments include any signal segment capable of directing the secretionof a protein of the present invention. Preferred signal segmentsinclude, but are not limited to, tissue plasminogen activator (t-PA),interferon, interleukin, growth hormone, histocompatibility and viralenvelope glycoprotein signal segments, as well as natural signalsegments. Suitable fusion segments encoded by fusion segment nucleicacids are disclosed herein. In addition, a nucleic acid molecule of thepresent invention can be joined to a fusion segment that directs theencoded protein to the proteosome, such as a ubiquitin fusion segment.Recombinant molecules may also include intervening and/or untranslatedsequences surrounding and/or within the nucleic acid sequences ofnucleic acid molecules of the present invention.

[0082] The following table summarizes the Sequence Listing, forconvenience: Description of sequence SEQ ID NO DNA sequence whichencodes a portion of a IgE heavy chain 1 AA sequence of a portion of aIgE heavy chain 2 reverse DNA complement to 1 3 DNA sequence whichencodes the most preferred FcεR binding region of the IgE heavy chain 4AA sequence which is the most preferred FcεR binding region of the IgEheavy chain 5 reverse DNA complement to 3 6 DNA sequence of morepreferred FcεR binding region 7 AA sequence of 7 8 reverse DNAcomplement to 7 9 DNA sequence of preferred FcεR binding region 10 AAsequence of 10 11 reverse DNA complement to 8 12 DNA sequence ofconstant region 13 AA sequence of constant region 14 reverse DNAcomplement of 13 15 DNA sequence of partial variable region 16 AAsequence of partial variable region 17 reverse DNA complement of 16 18DNA sequence which encodes the IgE light chain 19 AA sequence which isthe IgE light chain 20 reverse DNA complement to 19 21 DNA sequence ofpolyadenylation signal 22 DNA nucleotides 7-732 of SEQ ID NO:19 23reverse DNA complement of 23 (nuc 223-948 of SEQ ID NO:21) 24 DNAnucleotides 67-732 of SEQ ID NO:19 25 AA 21-242 of SEQ ID NO:20 26 DNAnucleotides 223-888 of SEQ ID NO:21 27 DNA sequence which encodes aportion of a IgE heavy chain 28 AA sequence of a portion of a IgE heavychain 29 reverse DNA complement to 28 30 DNA nucleotides 1-1488 of 28 31Reverse complement of 31 32 DNA nucleotides 1-1293 of SEQ ID NO:13 33Reverse complement of 33 (nuc 126-1418 of SEQ ID NO:15) 34

[0083] One embodiment of the present invention includes a recombinantvector, which includes at least one isolated nucleic acid molecule ofthe present invention, inserted into any vector capable of deliveringthe nucleic acid molecule into a host cell. Such a vector containsheterologous nucleic acid sequences, that is nucleic acid sequences thatare not naturally found adjacent to nucleic acid molecules of thepresent invention and that preferably are derived from a species otherthan the species from which the nucleic acid molecule(s) are derived.The vector can be either RNA or DNA, either prokaryotic or eukaryotic,and typically is a virus or a plasmid. Recombinant vectors can be usedin the cloning, sequencing, and/or otherwise manipulation of feline IgEnucleic acid molecules of the present invention.

[0084] One type of recombinant vector, referred to herein as arecombinant molecule, comprises a nucleic acid molecule of the presentinvention operatively linked to an expression vector. The phraseoperatively linked refers to insertion of a nucleic acid molecule intoan expression vector in a manner such that the molecule is able to beexpressed when transformed into a host cell. As used herein, anexpression vector is a DNA or RNA vector that is capable of transforminga host cell and of effecting expression of a specified nucleic acidmolecule. Preferably, the expression vector is also capable ofreplicating within the host cell. Expression vectors can be eitherprokaryotic or eukaryotic, and are typically viruses or plasmids.Expression vectors of the present invention include any vectors thatfunction (i.e., direct gene expression) in recombinant cells of thepresent invention, including in bacterial, fungal, endoparasite, insect,other animal, and plant cells. Preferred expression vectors of thepresent invention can direct gene expression in bacterial, yeast, insectand mammalian cells and more preferably in the cell types disclosedherein.

[0085] In particular, expression vectors of the present inventioncontain regulatory sequences such as transcription control sequences,translation control sequences, origins of replication, and otherregulatory sequences that are compatible with the recombinant cell andthat control the expression of nucleic acid molecules of the presentinvention. In particular, recombinant molecules of the present inventioninclude transcription control sequences. Transcription control sequencesare sequences which control the initiation, elongation, and terminationof transcription. Particularly important transcription control sequencesare those which control transcription initiation, such as promoter,enhancer, operator and repressor sequences. Suitable transcriptioncontrol sequences include any transcription control sequence that canfunction in at least one of the recombinant cells of the presentinvention. A variety of such transcription control sequences are knownto those skilled in the art. Preferred transcription control sequencesinclude those which function in bacterial, yeast, insect and mammaliancells, such as, but not limited to, tac, lac, trp, trc, oxy-pro,omp/lpp, rrnB, bacteriophage lambda (such as lambda PL and lambda PR andfusions that include such promoters), bacteriophage T7, T7lac,bacteriophage T3, bacteriophage SP6, bacteriophage SP01,metallothionein, alpha-mating factor, Pichia alcohol oxidase, alphavirussubgenomic promoters (such as Sindbis virus subgenomic promoters),antibiotic resistance gene, baculovirus, Heliothis zea insect virus,vaccinia virus, herpesvirus, raccoon poxvirus, other poxvirus,adenovirus, cytomegalovirus (such as intermediate early promoters),simian virus 40, retrovirus, actin, retroviral long terminal repeat,Rous sarcoma virus, heat shock, phosphate and nitrate transcriptioncontrol sequences as well as other sequences capable of controlling geneexpression in prokaryotic or eukaryotic cells. Additional suitabletranscription control sequences include tissue-specific promoters andenhancers as well as lymphokine-inducible promoters (e.g., promotersinducible by interferons or interleukins). Transcription controlsequences of the present invention can also include naturally occurringtranscription control sequences naturally associated with cats. Thepresent invention also comprises expression vectors comprising a nucleicacid molecule described herein.

[0086] Recombinant DNA technologies can be used to improve expression oftransformed nucleic acid molecules by manipulating, for example, thenumber of copies of the nucleic acid molecules within a host cell, theefficiency with which those nucleic acid molecules are transcribed, theefficiency with which the resultant transcripts are translated, and theefficiency of post-translational modifications. Recombinant techniquesuseful for increasing the expression of nucleic acid molecules of thepresent invention include, but are not limited to, operatively linkingnucleic acid molecules to high-copy number plasmids, integration of thenucleic acid molecules into one or more host cell chromosomes, additionof vector stability sequences to plasmids, substitutions ormodifications of transcription control signals (e.g., promoters,operators, enhancers), substitutions or modifications of translationalcontrol signals (e.g., ribosome binding sites, Shine-Dalgarnosequences), modification of nucleic acid molecules of the presentinvention to correspond to the codon usage of the host cell, deletion ofsequences that destabilize transcripts, and use of control signals thattemporally separate recombinant cell growth from recombinant enzymeproduction during fermentation. The activity of an expressed recombinantprotein of the present invention may be improved by fragmenting,modifying, or derivatizing nucleic acid molecules encoding such aprotein.

[0087] Also provided by the present invention are recombinant cellstransformed with a nucleic acid described herein.

[0088] Transformation of a nucleic acid molecule into a cell can beaccomplished by any method by which a nucleic acid molecule can beinserted into the cell. Transformation techniques include, but are notlimited to, transfection, electroporation, microinjection, lipofection,adsorption, and protoplast fusion. A recombinant cell may remainunicellular or may grow into a tissue, organ or a multicellularorganism. Transformed nucleic acid molecules of the present inventioncan remain extrachromosomal or can integrate into one or more siteswithin a chromosome of the transformed (i.e., recombinant) cell in sucha manner that their ability to be expressed is retained.

[0089] Suitable host cells to transform include any cell that can betransformed with a nucleic acid molecule of the present invention. Hostcells can be either untransformed cells or cells that are alreadytransformed with at least one nucleic acid molecule (e.g., nucleic acidmolecules encoding one or more proteins of the present invention and/orother proteins useful in the production of multivalent vaccines). Hostcells of the present invention either can be endogenously (i.e.,naturally) capable of producing feline IgE proteins of the presentinvention or can be capable of producing such proteins after beingtransformed with at least one nucleic acid molecule of the presentinvention. Host cells of the present invention can be any cell capableof producing at least one protein of the present invention, and includebacterial, fungal (including yeast), other insect, other animal andplant cells. Preferred host cells include bacterial, mycobacterial,yeast, parasite, insect and mammalian cells. More preferred host cellsinclude Salmonella, Escherichia, Bacillus, Listeria, Saccharomyces,Spodoptera, Mycobacteria, Trichoplusia, BHK (baby hamster kidney) cells,MDCK cells (normal dog kidney cell line for canine herpesviruscultivation), CRFK cells (normal cat kidney cell line for felineherpesvirus cultivation), CV-1 cells (African monkey kidney cell lineused, for example, to culture raccoon poxvirus), COS (e.g., COS-7)cells, and Vero cells. Particularly preferred host cells are Escherichiacoli, including E. coli K-12 derivatives; Salmonella typhi; Salmonellatyphimurium, including attenuated strains such as UK-1 _(X)3987 andSR-11 _(X)4072; Spodoptera frugiperda; Trichoplusia ni; BHK cells; MDCKcells; CRFK cells; CV-1 cells; COS cells; Vero cells; andnon-tumorigenic mouse myoblast G8 cells (e.g., ATCC CRL 1246).Additional appropriate mammalian cell hosts include other kidney celllines, other fibroblast cell lines (e.g., human, murine or chickenembryo fibroblast cell lines), myeloma cell lines, Chinese hamster ovarycells, mouse NIH/3T3 cells, LMTK³¹ cells and/or HeLa cells. In oneembodiment, the proteins may be expressed as heterologous proteins inmyeloma cell lines employing immunoglobulin promoters.

[0090] A recombinant cell is preferably produced by transforming a hostcell with one or more recombinant molecules, each comprising one or morenucleic acid molecules of the present invention operatively linked to anexpression vector containing one or more transcription controlsequences. The phrase “operatively linked” refers to insertion of anucleic acid molecule into an expression vector in a manner such thatthe molecule is able to be expressed when transformed into a host cell.

[0091] A recombinant molecule of the present invention is a moleculethat can include at least one of any nucleic acid molecule heretoforedescribed operatively linked to at least one of any transcriptioncontrol sequence capable of effectively regulating expression of thenucleic acid molecule(s) in the cell to be transformed, examples ofwhich are disclosed herein.

[0092] A recombinant cell of the present invention includes any celltransformed with at least one of any nucleic acid molecule of thepresent invention. Suitable and preferred nucleic acid molecules as wellas suitable and preferred recombinant molecules with which to transformcells are disclosed herein.

[0093] The present invention also provides isolated proteins encoding aportion of a feline IgE heavy chain molecule, wherein said proteinscomprise an amino acid sequence selected from the group consisting of:

[0094] (a) an amino acid sequence encoded by a nucleic acid sequencewhich has more than 82% identity to a nucleic acid sequence selectedfrom the group consisting of: SEQ ID NO 1; and SEQ ID NO: 28, whereinsaid identity can be determined using the DNAsis computer program anddefault parameters;

[0095] (b) an amino acid sequence which has more than 76% identity to anamino acid sequence selected from the group consisting of: SEQ ID NO 2;and SEQ ID NO: 29, wherein said identity can be determined using theDNAsis computer program and default parameters;

[0096] (c) an amino acid sequence encoded by an allelic variant of anucleic acid sequence selected from the group consisting of: SEQ ID NO1; and SEQ ID NO: 28; and

[0097] (d) an amino acid sequence encoded by a a nucleic acid sequencewhich has more than 90% identity to a nucleic acid sequence selectedfrom the group consisting of: SEQ ID NO 3; SEQ ID NO 4; SEQ ID NO 6; SEQID NO 7; SEQ ID NO 9; SEQ ID NO 10; SEQ ID NO 12; SEQ ID NO 13; SEQ IDNO 15; SEQ ID NO 16; SEQ ID NO 18; and SEQ ID NO 31.

[0098] Comparison of amino acid sequence SEQ ID NO 2 (i.e., the aminoacid sequence of the heavy chain of feline IgE) with amino acidsequences reported in GenBank™ indicates that SEQ ID NO 2 showed themost homology, i.e., about 76% identity, with IgE protein of Canisfamiliaris (GenBank accession number 598109).

[0099] Also provided are isolated proteins encoding a portion of afeline IgE light chain molecule, wherein said protein comprises an aminoacid sequence selected from the group consisting of:

[0100] (a) an amino acid sequence encoded by a nucleic acid sequencewhich has more than 84% identity to SEQ ID NO 19, and wherein saididentity can be determined using the DNAsis computer program and defaultparameters;

[0101] (b) an amino acid sequence which has more than 61% identity toSEQ ID NO 20, wherein said identity can be determined using the DNAsiscomputer program and default parameters;

[0102] (c) an amino acid sequence encoded by an allelic variant of SEQID NO 19; and

[0103] (d) a nucleic acid sequence which has more than 95% identity to anucleic acid sequence selected from the group consisting of: SEQ ID NO23; and SEQ ID NO 25. In another embodiment, there are provided isolatedfeline IgE light chain proteins, preferably, SEQ ID NO 19.

[0104] There are also provided recombinant cells comprising the proteinsherein described.

[0105] According to the present invention, a feline IgE protein of thepresent invention refers to: a feline IgE protein; a feline IgE homolog;a feline IgE mimetope; a feline IgE substrate analog; or a feline IgEpeptide. Preferably, a feline IgE molecule binds to Fcε receptors.

[0106] The present invention therefore provides proteins of the felineIgE. Both the light and heavy chains are provided, as are compositionscomprising the two, as well as portions of either. In particular,isolated feline constant region proteins are preferred, although Fcεbinding region proteins are most preferred. Proteins which would resultfrom expression of the nucleic acid molecules herein disclosed arepreferred, with the proteins which would result from expression of theexemplified compounds being most preferred. It is understood thatproteins which would result from expression of allelic variants of theexemplified sequences, as well as proteins which would result from theexpression of nucleic acid molecules which hybridize under stringenthybridization conditions to the nucleic acid molecules exemplified arewithin the scope of the present invention as well. Lastly, an amino acidsequence substantially homologous to a referent IgE protein will have atleast 85% sequence identity, preferably 90%, and most preferably 95%sequence homology with the amino acid sequence of a referent IgE proteinor a peptide thereof. For example, an amino acid sequence issubstantially homologous to feline IgE protein if, when aligned withfeline IgE protein, at least 85% of its amino acid residues are thesame. SEQ ID NO 2 and SEQ ID NO 29 are the most preferred proteins.

[0107] In another embodiment, a preferred feline IgE protein includes aprotein encoded by a nucleic acid molecule which is at least about 50nucleotides, or about 150 nucleotides, and which hybridizes underconditions which preferably allow about 35% base pair mismatch, morepreferably under conditions which allow about 30% base pair mismatch,more preferably under conditions which allow about 25% base pairmismatch, more preferably under conditions which allow about 20% basepair mismatch, more preferably under conditions which allow about 15%base pair mismatch, more preferably under conditions which allow about10% base pair mismatch, and even more preferably under conditions whichallow about 5% base pair mismatch with a nucleic acid molecule selectedfrom the group consisting of: SEQ ID NO 2; SEQ ID NO 5; SEQ ID NO 8; SEQID NO 11; SEQ ID NO 14; and SEQ ID NO 17.

[0108] Another embodiment of the present invention includes a feline IgEprotein encoded by a nucleic acid molecule selected from the groupconsisting of: a nucleic acid molecule comprising at least about 150nucleotides, wherein said nucleic acid molecule comprising at leastabout 150 nucleotides hybridizes, in a solution comprising 1×SSC and 0%formamide, at a temperature of about 56° C., to a nucleic acid sequenceselected from the group consisting of SEQ ID NO 1, SEQ ID NO 4, SEQ IDNO 7, SEQ ID NO 10, SEQ ID NO 13, SEQ ID NO 16, and/or SEQ ID NO 19; anda nucleic acid molecule comprising a fragment of any of said nucleicacid molecules comprising at least about 150 nucleotides.

[0109] Yet another preferred feline IgE protein of the present inventionincludes a protein encoded by a nucleic acid molecule which ispreferably about 45% identical, more preferably about 50% identical,more preferably about 55% identical, more preferably about 60%identical, more preferably about 65% identical, more preferably about70% identical, more preferably about 75% identical, more preferablyabout 80% identical, more preferably about 85% identical, morepreferably about 90% identical and even more preferably about 95%identical to a nucleic acid molecule having the nucleic acid sequenceSEQ ID NO 1, SEQ ID NO 4, SEQ ID NO 7, SEQ ID NO 10, SEQ ID NO 13, SEQID NO 16, and/or SEQ ID NO 19, and/or fragments of such proteins.Percent identity as used herein is determined using the Compare functionby maximum matching within the program DNAsis Version 2.1 using defaultparameters.

[0110] More preferred feline IgE proteins of the present inventioninclude proteins comprising amino acid sequences that are at least about50%, preferably at least about 55%, more preferably at least about 60%,even more preferably at least about 65%, even more preferably at leastabout 70%, even more preferably at least about 75%, even more preferablyat least about 80%, even more preferably at least about 85%, even morepreferably at least about 90%, and even more preferably at least about95%, identical to amino acid sequence SEQ ID NO 1, SEQ ID NO 4, SEQ IDNO 7, SEQ ID NO 10, SEQ ID NO 13, SEQ ID NO 16, and/or SEQ ID NO 19.

[0111] Preferred feline IgE proteins of the present invention includeproteins that are at least about 50%, preferably at least about 55%,more preferably at least about 60%, even more preferably at least about65%, even more preferably at least about 70%, even more preferably atleast 75%, even more preferably at least about 80%, even more preferablyat least about 85%, even more preferably at least 90%, and even morepreferably at least about 95% identical to SEQ ID NO 2, SEQ ID NO 5, SEQID NO 7, SEQ ID NO 11, and/or SEQ ID NO 14.

[0112] A feline IgE heavy chain protein of the present invention,including a homolog, can be identified in a straight-forward manner bythe protein's ability to bind to FcpE receptor. Examples of feline IgEprotein homologs include feline IgE proteins in which amino acids havebeen deleted (e.g., a truncated version of the protein, such as apeptide), inserted, inverted, substituted and/or derivatized (e.g., byglycosylation, phosphorylation, acetylation, myristoylation,prenylation, palmitoylation, amidation and/or addition ofglycerophosphatidyl inositol) such that the homolog is capable ofbinding to Fcε receptor.

[0113] Feline IgE protein homologs can be the result of natural allelicvariation or natural mutation. Feline IgE protein homologs of thepresent invention can also be produced using techniques known in the artincluding, but not limited to, direct modifications to the protein ormodifications to the gene encoding the protein using, for example,classic or recombinant nucleic acid techniques to effect random ortargeted mutagenesis.

[0114] The minimal size of an IgE protein homolog of the presentinvention is a size sufficient to be encoded by a nucleic acid moleculecapable of forming a stable hybrid (i.e., hybridize under stringenthybridization conditions) with the complementary sequence of a nucleicacid molecule encoding the corresponding natural protein. As such, thesize of the nucleic acid molecule encoding such a protein homolog isdependent on nucleic acid composition and percent homology between thenucleic acid molecule and complementary sequence. It should also benoted that the extent of homology required to form a stable hybrid canvary depending on whether the homologous sequences are interspersedthroughout the nucleic acid molecules or are clustered (i.e., localized)in distinct regions on the nucleic acid molecules. The minimal size ofsuch nucleic acid molecules is typically at least about 12 to about 15nucleotides in length if the nucleic acid molecules are GC-rich and atleast about 15 to about 17 bases in length if they are AT-rich. As such,the minimal size of a nucleic acid molecule used to encode a feline IgEprotein homolog of the present invention is from about 12 to about 18nucleotides in length. Thus, the minimal size of a feline IgE proteinhomolog of the present invention is from about 4 to about 6 amino acidsin length. There is no limit, other than a practical limit, on themaximal size of such a nucleic acid molecule in that the nucleic acidmolecule can include a portion of a gene, an entire gene, multiplegenes, or portions thereof. The preferred size of a protein encoded by anucleic acid molecule of the present invention depends on whether afull-length, fusion, multivalent, or functional portion of such aprotein is desired. Preferably, the preferred size of a protein encodedby a nucleic acid molecule of the present invention is a portion of theprotein that binds to IgE which is about 30 amino acids, more preferablyabout 35 amino acids and even more preferably about 44 amino acids inlength.

[0115] As used herein, a feline refers to any member of the cat family,including domestic cats, wild cats and zoo cats. Examples of cats fromwhich to isolate feline IgE proteins of the present invention (includingisolation of the natural protein or production of the protein byrecombinant or synthetic techniques) include, but are not limited to,domestic cats, lions, tigers, leopards, panthers, cougars, bobcats,lynx, jaguars, cheetahs, and servals, with domestic cats being morepreferred and Felis domesticus cats being even more preferred.

[0116] The present invention also includes mimetopes of feline IgEproteins of the present invention. As used herein, a mimetope of afeline IgE protein of the present invention refers to any compound thatis able to mimic the activity of such a feline IgE protein (e.g.,ability to bind to Fcε receptors), often because the mimetope has astructure that mimics the feline IgE protein. It is to be noted,however, that the mimetope need not have a structure similar to a felineIgE protein as long as the mimetope functionally mimics the protein.Mimetopes can be, but are not limited to: peptides that have beenmodified to decrease their susceptibility to degradation; anti-idiotypicand/or catalytic antibodies, or fragments thereof; non-proteinaceousimmunogenic portions of an isolated protein (e.g., carbohydratestructures); synthetic or natural organic or inorganic molecules,including nucleic acids; and/or any other peptidomimetic compounds.Mimetopes of the present invention can be designed usingcomputer-generated structures of feline IgE proteins of the presentinvention. Mimetopes can also be obtained by generating random samplesof molecules, such as oligonucleotides, peptides or other organicmolecules, and screening such samples by affinity chromatographytechniques using the corresponding binding partner, (e.g., a feline Fcεreceptor domain or anti-feline IgE antibody). A mimetope can also beobtained by, for example, rational drug design. In a rational drugdesign procedure, the three-dimensional structure of a compound of thepresent invention can be analyzed by, for example, nuclear magneticresonance (NMR) or x-ray crystallography. The three-dimensionalstructure can then be used to predict structures of potential mimetopesby, for example, computer modeling. The predicted mimetope structurescan then be produced by, for example, chemical synthesis, recombinantDNA technology, or by isolating a mimetope from a natural source.Specific examples of feline IgE mimetopes include anti-idiotypicantibodies, oligonucleotides produced using Selex™ technology, peptidesidentified by random screening of peptide libraries and proteinsidentified by phage display technology. A preferred mimetope is apeptidomimetic compound that is structurally and/or functionally similarto a feline IgE protein of the present invention, particularly to theFcε receptor-binding domain of the feline IgE protein.

[0117] One embodiment of a feline IgE protein of the present inventionis a fusion protein that includes a feline IgE protein domain attachedto one or more fusion segments. Suitable fusion segments for use withthe present invention include, but are not limited to, segments thatcan: enhance a protein's stability; act as an immunopotentiator toenhance an immune response; act as an suppressor of immune responseand/or assist purification of a feline IgE protein (e.g., by affinitychromatography). A suitable fusion segment can be a domain of any sizethat has the desired function (e.g., imparts increased stability,imparts increased immunogenicity to a protein, and/or simplifiespurification of a protein). Fusion segments can be joined to aminoand/or carboxyl termini of the feline IgE-containing domain of theprotein and can be susceptible to cleavage in order to enablestraight-forward recovery of a feline IgE protein. Fusion proteins arepreferably produced by culturing a recombinant cell transformed with afusion nucleic acid molecule that encodes a protein including the fusionsegment attached to either the carboxyl and/or amino terminal end of afeline IgE-containing domain. Preferred fusion segments include a metalbinding domain (e.g., a poly-histidine segment); an immunoglobulinbinding domain (e.g., Protein A; Protein G; T cell; B cell; Fc receptoror complement protein antibody-binding domains); a sugar binding domain(e.g., a maltose binding domain); a “tag” domain (e.g., at least aportion of β-galactosidase, a strep tag peptide, other domains that canbe purified using compounds that bind to the domain, such as monoclonalantibodies); and/or a linker and enzyme domain (e.g., alkalinephosphatase domain connected to a feline IgE protein by a linker). Morepreferred fusion segments include metal binding domains, such as apoly-histidine segment; a maltose binding domain; a strep tag peptide,such as that available from Biometra in Tampa, Fla.; and a phage T7 S10peptide.

[0118] A feline IgE molecule of the present invention can also includechimeric molecules comprising a portion of a feline IgE molecule thatbinds to an Fcε receptor and a second molecule that enables the chimericmolecule to be bound to a substrate in such a manner that the IgEmolecule portion binds to FcεR in essentially the same manner as an IgEmolecule that is not bound to a substrate. An example of a suitablesecond molecule includes a portion of an immunoglobulin molecule oranother ligand that has a suitable binding partner that can beimmobilized on a substrate, e.g., biotin and avidin, or a metal-bindingprotein and a metal (e.g., His), or a sugar-binding protein and a sugar(e.g., maltose).

[0119] Chimeric immunoglobulin molecules are also included in thepresent invention. Specifically, a chimeric immunoglobulin moleculewhich contains a portion from a feline IgE and a portion that is notfeline is contemplated. The non-feline portion is ideally the antigenbinding site of the IgE, and therefore, should include less than about1% non-feline sequence. A chimeric molecule ideally contains only thoseportions of the non-feline variable region that binds to antigen, withthe remainder of the immunoglobulin comprising feline sequence.

[0120] A variety of procedures known in the art may be used tomolecularly clone feline IgE DNA of the present invention. These methodsinclude, but are not limited to, direct functional expression of thefeline IgE genes following the construction of feline IgE-containingcDNA or genomic DNA library in an appropriate expression vector system.Another method is to screen feline IgE-containing cDNA or genomic DNAlibrary constructed in a bacteriophage or plasmid shuttle vector with alabeled oligonucleotide probe designed from the amino acid sequence ofthe feline IgE subunits. An additional method consists of screening afeline IgE-containing cDNA or genomic DNA libraries constructed in abacteriophage or plasmid shuttle vector with a partial DNA encoding thefeline IgE. This partial DNA is obtained by the specific PCRamplification of feline IgE DNA fragments through the design ofdegenerate oligonucleotide primers from the amino acid sequence of thepurified feline IgE.

[0121] The translation of the RNA into a peptide or a protein willresult in the production of at least a portion of the feline IgE proteinwhich can be identified, for example, by the activity of feline IgEprotein or by immunological reactivity with an anti-feline IgE antibody.In this method, pools of mRNA isolated from feline IgE-producing cellscan be analyzed for the presence of an RNA which encodes at least aportion of the feline IgE protein. Further fractionation of the RNA poolcan be done to purify the feline IgE RNA from non-feline IgE RNA. Thepeptide or protein produced by this method may be analyzed to provideamino acid sequences which in turn are used to provide primers forproduction of feline IgE cDNA, or the RNA used for translation can beanalyzed to provide nucleotide sequences encoding feline IgE and produceprobes for the production of feline IgE cDNA. These methods are known inthe art and can be found in, for example, Sambrook, J., Fritsch, E. F.,Maniatis, T. in Molecular Cloning: A Laboratory Manual, Second Edition,Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. 1989.

[0122] Other types of libraries, as well as libraries constructed fromother cells or cell types, may be useful for isolating felineIgE-encoding DNA. Other types of libraries include, but are not limitedto, cDNA libraries derived from other felines or cell lines derived fromother felines, and genomic DNA libraries. Preparation of cDNA librariescan be performed by standard techniques. Well known cDNA libraryconstruction techniques can be found in, for example, Sambrook, J., etal., ibid.

[0123] DNA encoding feline IgE may also be isolated from a suitablegenomic DNA library. Construction of genomic DNA libraries can beperformed by standard techniques. Well known genomic DNA libraryconstruction techniques can be found in Sambrook, J., et al., ibid.

[0124] In order to clone the feline IgE gene by the above methods,knowledge of the amino acid sequence of feline IgE may be necessary. Onemay either use the sequences herein exemplified or purify feline IgEprotein and sequence a portion of the protein by manual or automatedsequencing. It is not necessary to determine the entire amino acidsequence, because the linear sequence of two regions of 6 to 8 aminoacids from the protein can be determined and used to produce primers forPCR amplification of feline IgE DNA.

[0125] Once suitable amino acid sequences have been identified, the DNAsequences capable of encoding them are synthesized. Because the geneticcode is degenerate, more than one codon may be used to encode aparticular amino acid, and therefore, the amino acid sequence can beencoded by any of a set of similar DNA oligonucleotides. Only one memberof the set will be identical to the feline IgE sequence but will becapable of hybridizing to feline IgE DNA even in the presence of DNAoligonucleotides with mismatches under appropriate conditions. Underalternate conditions, the mismatched DNA oligonucleotides may stillsufficiently hybridize to the feline IgE DNA to permit identificationand isolation of feline IgE encoding DNA.

[0126] In one embodiment, an isolated protein of the present inventionis produced by culturing a cell capable of expressing the protein underconditions effective to produce the protein, and recovering the protein.A preferred cell to culture is a recombinant cell of the presentinvention. Effective culture conditions include, but are not limited to,effective media, bioreactor, temperature, pH and oxygen conditions thatpermit protein production. An effective medium refers to any medium inwhich a cell is cultured to produce a feline IgE protein of the presentinvention. Such a medium typically comprises an aqueous medium havingassimilable carbon, nitrogen and phosphate sources, and appropriatesalts, minerals, metals and other nutrients, such as vitamins. Cells ofthe present invention can be cultured in conventional fermentationbioreactors, shake flasks, test tubes, microtiter dishes, and petriplates. Culturing can be carried out at a temperature, pH and oxygencontent appropriate for a recombinant cell. Such culturing conditionsare within the expertise of one of ordinary skill in the art.

[0127] Depending on the vector and host system used for production,resultant proteins of the present invention may either remain within therecombinant cell; be secreted into the fermentation medium; be secretedinto a space between two cellular membranes, such as the periplasmicspace in E. coli; or be retained on the outer surface of a cell or viralmembrane. The phrase “recovering the protein”, as well as similarphrases, refers to collecting the whole fermentation medium containingthe protein and need not imply additional steps of separation orpurification. Proteins of the present invention can be purified using avariety of standard protein purification techniques, such as, but notlimited to, affinity chromatography, ion exchange chromatography,filtration, electrophoresis, hydrophobic interaction chromatography, gelfiltration chromatography, reverse phase chromatography, concanavalin Achromatography, chromatofocusing and differential solubilization.Proteins of the present invention are preferably retrieved in“substantially pure” form. As used herein, “substantially pure” refersto a purity that allows for the effective use of the protein as atherapeutic composition or diagnostic. A therapeutic composition foranimals, for example, should exhibit few impurities.

[0128] In addition, recombinant feline IgE can be separated from othercellular proteins by use of an immunoaffinity column made withmonoclonal or polyclonal antibodies specific for full length nascentfeline IgE, or polypeptide fragments of feline IgE.

[0129] The present invention also includes isolated (i.e., removed fromtheir natural milieu) antibodies that selectively bind to a feline IgEprotein of the present invention or a mimetope thereof (i.e.,anti-feline IgE antibodies). As used herein, the term “selectively bindsto” a feline IgE protein refers to the ability of antibodies of thepresent invention to preferentially bind to specified proteins andmimetopes thereof of the present invention. Binding can be measuredusing a variety of methods standard in the art including enzymeimmunoassays (e.g., ELISA), immunoblot assays, etc.; see, for example,Sambrook et al., ibid. An anti-feline IgE antibody preferablyselectively binds to a feline IgE protein in such a way as to reduce theactivity of that protein.

[0130] In particular, there are provided antibodies directed to thefeline IgE. In particular, antibodies that bind specifically to theheavy and/or light chain of IgE are provided. Preferred are antibodiesselective for the constant region of the feline IgE heavy chain,although more preferred are antibodies selective for the Fcεreceptor-binding domain of the IgE heavy chain. In one preferredembodiment, there are provided antibodies selective for a proteinselected from the group consisting of: SEQ ID NO 2; SEQ ID NO 5; SEQ IDNO 8; SEQ ID NO 11; SEQ ID NO 14; SEQ ID NO 17; SEQ ID NO 20; SEQ ID NO26; and SEQ ID NO 29. These antibodies may be admixed or conjugated withadditional materials, such as cytotic agents or other antibodyfragments, including IgG fragments. In particular, antibodies asdescribed in the examples are included, and preferred embodiments of thepresent invention, such as, H-100, H-101, H-102, H-103, H-106. However,those antibodies specific for the feline IgE light chain are alsoincluded, especially: H-99, H-104, and H-107.

[0131] Isolated antibodies of the present invention can includeantibodies in a bodily fluid (such as, but not limited to, serum), orantibodies that have been purified to varying degrees. Antibodies of thepresent invention can be polyclonal or monoclonal. Functionalequivalents of such antibodies, such as antibody fragments andgenetically-engineered antibodies (including single chain antibodies orchimeric antibodies that can bind to more than one epitope) are alsoincluded in the present invention.

[0132] A preferred method to produce antibodies of the present inventionincludes (a) administering to an animal an effective amount of aprotein, peptide or mimetope thereof of the present invention to producethe antibodies and (b) recovering the antibodies. In another method,antibodies of the present invention are produced recombinantly usingtechniques as heretofore disclosed to produce feline IgE proteins of thepresent invention. Antibodies raised against defined proteins ormimetopes can be advantageous because such antibodies are notsubstantially contaminated with antibodies against other substances thatmight otherwise cause interference in a diagnostic assay or side effectsif used in a therapeutic composition.

[0133] Antibodies of the present invention have a variety of potentialuses that are within the scope of the present invention. For example,such antibodies can be used (a) as tools to detect IgE in the presenceor absence of Fcε receptor and/or (b) as tools to screen expressionlibraries and/or to recover desired proteins of the present inventionfrom a mixture of proteins and other contaminants. Furthermore,antibodies of the present invention can be used to target cytotoxicagents to cells having Fcε receptors in order to directly kill suchcells. Targeting can be accomplished by conjugating (i.e., stablyjoining) such antibodies to the cytotoxic agents using techniques knownto those skilled in the art. Suitable cytotoxic agents are known tothose skilled in the art. Antibodies of the present invention, includingFcε receptor binding site-binding portions thereof, can also be used,for example, to inhibit binding of IgE to Fcε receptors, to produceanti-feline IgE idiotypic antibodies, to purify cells having feline IgEproteins, to stimulate intracellular signal transduction through afeline Fcε and to identify cells having feline IgE proteins.

[0134] The above described methods for producing monospecific antibodiesmay be utilized to produce antibodies specific for feline IgEpolypeptide fragments, or full-length nascent feline IgE polypeptide.

[0135] Polyclonal serum may be obtained by well-known methods, such asby immunizing an animal (ie. rabbit), with a feline IgE and isolatingserum.

[0136] Another embodiment of the present invention are therapeuticcompositions that, when administered to an animal in an effectivemanner, are capable of affecting IgE-mediated reactions associated withdiseases related to biological responses involving IgE function. Atherapeutic composition of the present invention can include: a nucleicacid of the present invention, a protein of the present invention or aninhibitor of the present invention.

[0137] By “inhibitor” it is meant that the compound inhibits theformation of a complex between feline IgE protein and Fcε receptor. Suchinhibitors can, for example, interact with the feline Fcε receptorbinding site on IgE, other regions on feline IgE that effect IgE bindingto Fcε, receptor or the IgE binding site, for example, by allostericinteraction, on Fcε receptor. An inhibitor of IgE and Fcε receptorcomplex formation protein can interfere with complex formation by, forexample, preventing formation of an IgE protein and Fcε receptor complexor disrupting an existing IgE protein and Fcε receptor complex causingthe IgE protein and Fcε receptor to dissociate. An inhibitor of IgE andFcε receptor complex formation is usually a relatively small molecule.Preferably, an inhibitor of the present invention is derived from afeline IgE of the present invention, and more preferably from the Fcεreceptor binding site of the IgE, and is identified by its ability tobind to, or otherwise interact with, a Fcε receptor protein, therebyinterfering with the formation of a complex between a feline IgE proteinand Fcε receptor.

[0138] Preferred inhibitors of a feline IgE protein of the presentinvention include, but are not limited to, feline IgE proteins,fragments or mimetopes thereof, a Fcε receptor binding analog of afeline IgE protein, and other molecules that bind to a feline IgEprotein (e.g., to an allosteric site) or Fcε receptor in such a mannerthat Fcε receptor and IgE protein complex formation is inhibited.Preferred feline IgE proteins, fragments and mimetopes thereof arecapable of binding to Fcε receptor in such a manner that feline IgE doesnot bind to Fcε receptor. Mimetopes include those disclosed herein.

[0139] A feline IgE protein binding analog refers to a compound thatinteracts with (e.g., binds to, associates with, modifies) the Fcεreceptor-binding site of a feline lgE protein. A preferred feline IgEprotein binding analog inhibits Fcε receptor-binding activity of afeline IgE protein. Feline IgE protein binding analogs can be of anyinorganic or organic composition, and, as such, can be, but are notlimited to, peptides, nucleic acids, and peptidomimetic compounds.Feline IgE protein substrate analogs can be, but need not be,structurally similar to a feline IgE protein's natural substrate (e.g.,Fcε receptor) as long as they can interact with the active site (e.g.,Fcε receptor-binding site of that feline IgE). Feline IgE proteinbinding analogs can be designed using computer-generated structures offeline IgE proteins of the present invention or computer structures of,for example, the IgE-binding domain of Fcε receptor. Binding analogs canalso be obtained by generating random samples of molecules, such asoligonucleotides, peptides, peptidomimetic compounds, or other inorganicor organic molecules, and screening such samples by affinitychromatography techniques using the corresponding binding partner,(e.g., a feline IgE protein or anti-feline IgE idiotypic antibody). Apreferred feline IgE protein binding analog is a peptidomimetic compound(i.e., a compound that is structurally and/or functionally similar to anatural feline Fcε, receptor protein, particularly to the region of thesubstrate that binds to a feline IgE protein, but that inhibits IgEbinding upon interacting with the Fcε receptor binding site).

[0140] Feline IgE molecules, as well as other inhibitors and therapeuticcompounds, can be used directly as compounds in compositions of thepresent invention to treat animals as long as such compounds are notharmful to the animals being treated.

[0141] The present invention also includes a therapeutic compositioncomprising one or more therapeutic compounds of the present invention.Examples of such therapeutic compounds are disclosed herein.

[0142] A therapeutic composition of the present invention can be used toreduce an IgE-mediated biological response in an animal by administeringsuch a composition to an animal. Preferably, an animal is treated byadministering to the animal a therapeutic composition of the presentinvention in such a manner that a therapeutic compound (e.g., aninhibitor of a feline IgE protein, an anti-feline IgE antibody, aninhibitor of Fcε receptor, or nucleic acid molecules encoding feline IgEproteins) binds to an IgE molecule in the animal. Such administrationcould be by a variety of routes known to those skilled in the artincluding, but not limited to, subcutaneous, intradermal, intravenous,intranasal, oral, transdermal, intramuscular routes and other parenteralroutes.

[0143] Compositions of the present invention can be administered to anyanimal having an IgE that binds to a therapeutic compound of the presentinvention or to a protein expressed by a nucleic acid molecule containedin a therapeutic composition. Preferred animals to treat include mammalsand birds, with cats, dogs, horses, humans and other pets, work and/oreconomic food animals. Particularly preferred animals to protect arecats and dogs.

[0144] Therapeutic compositions of the present invention can beformulated in an excipient that the animal to be treated can tolerate.Examples of such excipients include water, saline, Ringer's solution,dextrose solution, Hank's solution, and other aqueous physiologicallybalanced salt solutions. Nonaqueous vehicles, such as fixed oils, sesameoil, ethyl oleate, or triglycerides may also be used. Other usefulformulations include suspensions containing viscosity enhancing agents,such as sodium carboxymethylcellulose, sorbitol, or dextran. Excipientscan also contain minor amounts of additives, such as substances thatenhance isotonicity and chemical stability. Examples of buffers includephosphate buffer, bicarbonate buffer and Tris buffer, while examples ofpreservatives include thimerosal, cresols, formalin and benzyl alcohol.Standard formulations can either be liquid injectables or solids whichcan be taken up in a suitable liquid as a suspension or solution forinjection. Thus, in a non-liquid formulation, the excipient can comprisedextrose, human serum albumin, preservatives, etc., to which sterilewater or saline can be added prior to administration.

[0145] In one embodiment of the present invention, a therapeuticcomposition can include an adjuvant. Adjuvants are agents that arecapable of enhancing the immune response of an animal to a specificantigen. Suitable adjuvants include, but are not limited to, cytokines,chemokines, and compounds that induce the production of cytokines andchemokines (e.g., granulocyte macrophage colony stimulating factor(GM-CSF), granulocyte colony stimulating factor (G-CSF), macrophagecolony stimulating factor (M-CSF), colony stimulating factor (CSF),Flt-3 ligand, erythropoietin (EPO), interleukin 2 (IL-2), interleukin-3(IL-3), interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6(IL-6), interleukin 7 (IL-7), interleukin 8 (IL-8), interleukin 10(IL-10), interleukin 12 (IL-12), interferon gamma, interferon gammainducing factor I (IGIF), transforming growth factor beta, RANTES(regulated upon activation, normal T cell expressed and presumablysecreted), macrophage inflammatory proteins (e.g., MIP-1 alpha and MIP-1beta), and Leishmania elongation initiating factor (LEIF); bacterialcomponents (e.g., endotoxins, in particular superantigens, exotoxins andcell wall components); aluminum-based salts; calcium-based salts;silica; polynucleotides; toxoids; serum proteins, viral coat proteins;block copolymer adjuvants (e.g., Hunter's Titermax™ adjuvant (Vaxcel™,Inc. Norcross, Ga.), Ribi adjuvants (Ribi ImmunoChem Research, Inc.,Hamilton, Mont.); and saponins and their derivatives (e.g., Quil A(Superfos Biosector A/S, Denmark). Protein adjuvants of the presentinvention can be delivered in the form of the protein themselves or ofnucleic acid molecules encoding such proteins using the methodsdescribed herein.

[0146] In one embodiment of the present invention, a therapeuticcomposition can include a carrier. Carriers include compounds thatincrease the half-life of a therapeutic composition in the treatedanimal. Suitable carriers include, but are not limited to, polymericcontrolled release vehicles, biodegradable implants, liposomes,bacteria, viruses, other cells, oils, esters, and glycols.

[0147] One embodiment of the present invention is a controlled releaseformulation that is capable of slowly releasing a composition of thepresent invention into an animal. As used herein, a controlled releaseformulation comprises a composition of the present invention in acontrolled release vehicle. Suitable controlled release vehiclesinclude, but are not limited to, biocompatible polymers, other polymericmatrices, capsules, microcapsules, microparticles, bolus preparations,osmotic pumps, diffusion devices, liposomes, lipospheres, andtransdermal delivery systems. Other controlled release formulations ofthe present invention include liquids that, upon administration to ananimal, form a solid or a gel in situ. Preferred controlled releaseformulations are biodegradable (i.e., bioerodible).

[0148] A preferred controlled release formulation of the presentinvention is capable of releasing a composition of the present inventioninto the blood of an animal at a constant rate sufficient to attaintherapeutic dose levels of the composition to reduce Fcε,receptor-mediated biological responses in the animal. As used herein,Fcε receptor-mediated biological response refers to cellular responsesthat occur when IgE is complexed with Fcε receptor. For example, aFcε-mediated biological response includes release of biologicalmediators, such as histamine, prostaglandins and/or proteases, that cantrigger clinical symptoms of allergy. The therapeutic composition ispreferably released over a period of time ranging from about 1 to about12 months. A preferred controlled release formulation of the presentinvention is capable of effecting a treatment preferably for at leastabout 1 month, more preferably for at least about 3 months, even morepreferably for at least about 6 months, even more preferably for atleast about 9 months, and even more preferably for at least about 12months.

[0149] Acceptable protocols to administer therapeutic compositions ofthe present invention in an effective manner include individual dosesize, number of doses, frequency of dose administration, and mode ofadministration. Determination of such protocols can be accomplished bythose skilled in the art. A suitable single dose is a dose that iscapable of protecting (i.e., preventing or treating) an animal fromdisease when administered one or more times over a suitable time period.The need for additional administrations of a therapeutic composition canbe determined by one of skill in the art in accordance with the givencondition of a patient. For example, to regulate an antigen-specific Fcεreceptor-mediated response, a therapeutic composition may beadministered more frequently when an antigen is present in a patient'senvironment in high amounts and less frequently when the antigen ispresent in lower amounts.

[0150] According to one embodiment, a nucleic acid molecule of thepresent invention can be administered to an animal in a fashion toenable expression of that nucleic acid molecule into a feline IgEprotein or a feline IgE RNA (e.g., antisense RNA, ribozyme, triple helixforms or RNA drug) in the animal. Nucleic acid molecules can bedelivered to an animal in a variety of methods including, but notlimited to, (a) administering a naked (i.e., not packaged in a viralcoat or cellular membrane) nucleic acid molecule (e.g., as naked DNA orRNA molecules, such as is taught, for example in Wolff et al., 1990,Science 247, 1465-1468) or (b) administering a nucleic acid moleculepackaged as a recombinant virus or as a recombinant cell (i.e., thenucleic acid molecule is delivered by a viral or cellular vehicle).

[0151] A naked nucleic acid molecule of the present invention includes anucleic acid molecule of the present invention and preferably includes arecombinant molecule of the present invention that preferably isreplication, or otherwise amplification, competent. A naked nucleic acidof the present invention can comprise one or more nucleic acid moleculesof the present invention in the form of, for example, a bicistronicrecombinant molecule having, for example one or more internal ribosomeentry sites. Preferred naked nucleic acid molecules include at least aportion of a viral genome (i.e., a viral vector). Preferred viralvectors include those based on alphaviruses, poxviruses, adenoviruses,herpesviruses, picornaviruses, and retroviruses, with those based onalphaviruses (such as Sindbis or Semliki virus), species-specificherpesviruses and species-specific poxviruses being particularlypreferred. Any suitable transcription control sequence can be used,including those disclosed as suitable for protein production.Particularly preferred transcription control sequence includecytomegalovirus intermediate early (preferably in conjunction withIntron-A), Rous Sarcoma Virus long terminal repeat, and tissue-specifictranscription control sequences, as well as transcription controlsequences endogenous to viral vectors if viral vectors are used. Theincorporation of “strong” poly(A) sequences are also preferred.

[0152] Naked nucleic acid molecules of the present invention can beadministered by a variety of methods. Suitable delivery methods include,for example, intramuscular injection, subcutaneous injection,intradermal injection, intradermal scarification, particle bombardment,oral application, and nasal application, with intramuscular injection,intradermal injection, intradermal scarification and particlebombardment being preferred. A preferred single dose of a naked DNAmolecule ranges from about 1 nanogram (ng) to about 1 milligram (mg),depending on the route of administration and/or method of delivery, ascan be determined by those skilled in the art. Examples ofadministration methods are disclosed, for example, in U.S. Pat. No.5,204,253, by Bruner, et al., issued Apr. 20, 1993, PCT Publication No.W0 95/19799, published Jul. 27, 1995, by McCabe, and PCT Publication No.WO 95/05853, published Mar. 2, 1995, by Carson, et al. Naked DNAmolecules of the present invention can be contained in an aqueousexcipient (e.g., phosphate buffered saline) and/or with a carrier (e.g.,lipid-based vehicles), or it can be bound to microparticles (e.g., goldparticles).

[0153] A recombinant virus of the present invention includes arecombinant molecule of the present invention that is packaged in aviral coat and that can be expressed in an animal after administration.Preferably, the recombinant molecule is packaging-deficient and/orencodes an attenuated virus. A number of recombinant viruses can beused, including, but not limited to, those based on alphaviruses,poxviruses, adenoviruses, herpesviruses, picornaviruses andretroviruses. Preferred recombinant viruses are those based onalphaviruses (such as Sindbis virus), raccoon poxviruses,species-specific herpesviruses and species-specific poxviruses. Anexample of methods to produce and use alphavirus recombinant virus isdisclosed in PCT Publication No. WO 94/17813, by Xiong et al., publishedAug. 18, 1994, which is incorporated by reference herein in itsentirety.

[0154] When administered to an animal, a recombinant virus of thepresent invention infects cells within the recipient animal and directsthe production of a protein or RNA nucleic acid molecule that is capableof reducing Fcε receptor-mediated biological responses in the animal.For example, a recombinant virus comprising a feline IgE nucleic acidmolecule of the present invention is administered according to aprotocol that results in the animal producing an amount of protein orRNA sufficient to reduce IgE-mediated biological responses. A preferredsingle dose of a recombinant virus of the present invention is fromabout 1×10⁴ to about 1×10⁷ virus plaque forming units (pfu) per kilogrambody weight of the animal. Administration protocols are similar to thosedescribed herein for protein-based compositions, with subcutaneous,intramuscular, intranasal and oral administration routes beingpreferred.

[0155] A recombinant cell useful in a therapeutic composition of thepresent invention includes recombinant cells of the present inventionthat comprises at least one feline IgE of the present invention.Preferred recombinant cells for this embodiment include Salmonella, E.coli, Listeria, Mycobacterium, S. frugiperda, yeast, (includingSaccharomyces cerevisiae), BHK, CV-1, myoblast G8, COS (e.g., COS-7),Vero, MDCK and CRFK recombinant cells. A recombinant cell of the presentinvention can be administered in a variety of ways but have theadvantage that they can be administered orally, preferably at dosesranging from about 10⁸ to about 10¹² cells per kilogram body weight.Administration protocols are similar to those described herein forprotein compositions. Recombinant cells can comprise whole cells, cellsstripped of cell walls or cell lysates. Pharmaceutically usefulcompositions comprising feline IgE DNA, feline IgE RNA, or feline IgEprotein, or other modulators of feline IgE activity, such as mimetopes,analogs, homologs, chimeras which inhibit the IgE/Fcε receptorinteraction, may be formulated according to known methods such as by theadmixture of a pharmaceutically acceptable carrier, or by modificationwith additional chemical moieties so as to form a chemical derivative.Examples of such carriers, modifications and methods of formulation maybe found in Remington's Pharmaceutical Sciences. To form apharmaceutically acceptable composition suitable for effectiveadministration, such compositions will contain an effective amount ofthe protein, DNA, RNA, or modulator.

[0156] The present invention also has the objective of providingsuitable topical, oral, systemic and parenteral formulations of thepharmaceutical compounds herein provided. The formulations can beadministered in a wide variety of therapeutic dosage forms inconventional vehicles for administration. For example, the compounds canbe formulated for oral administration in the form of tablets, capsules(each including timed release and sustained release formulations),pills, powders, granules, elixirs, tinctures, solutions, suspensions,syrups and emulsions, or by injection. Likewise, they may also beadministered in intravenous (both bolus and infusion), intraperitoneal,subcutaneous, topical with or without occlusion, or intramuscular form,all using forms well known to those of ordinary skill in thepharmaceutical arts. An effective but non-toxic amount of the compounddesired can be employed as a feline IgE modulating agent.

[0157] In addition, a feline IgE molecule formulation of the presentinvention can include not only a feline IgE molecule but also one ormore additional antigens or antibodies useful to affect immunogenicchange in an animal. As used herein, an antigen refers to any moleculecapable of being selectively bound by an antibody. As used herein,selective binding of a first molecule to a second molecule refers to theability of the first molecule to preferentially bind (e.g., havinghigher affinity higher avidity) to the second molecule when compared tothe ability of a first molecule to bind to a third molecule. The firstmolecule need not necessarily be the natural ligand of the secondmolecule. Examples of such antibodies include, but are not limited to,antibodies that bind selectively to the constant region of an IgE heavy(i.e., anti-IgE isotype antibody) or antibodies that bind selectively toan IgE having a specific antigen specificity (i.e., anti-IgE idiotypicantibody). Suitable anti-IgE antibodies for use in a formulation of thepresent invention are not capable of cross-linking two or more IgEantibodies. Preferred anti-IgE antibodies include Fab fragments of theantibodies (as defined in Janeway et al., ibid.). Examples of suchantigens include any antigen known to induce immunogenic change in ananimal. Preferred antigens include allergens and parasite antigens.Allergens of the present invention are preferably derived from fungi,trees, weeds, shrubs, grasses, wheat, corn, soybeans, rice, eggs, milk,cheese, bovines (or cattle), poultry, swine, cats, sheep, yeast, fleas,flies, mosquitos, mites, midges, biting gnats, lice, bees, wasps, ants,true bugs or ticks. A suitable flea allergen includes an allergenderived from a flea, in particular flea saliva antigen. A preferred fleaallergen includes a flea saliva antigen. Preferred flea saliva antigensinclude antigens such as those disclosed in PCT Patent Publication No.WO 96/11271, published Apr. 18, 1996, by Frank et al. (this publicationis incorporated by reference herein in its entirety), with flea salivaproducts and flea saliva proteins being particularly preferred.According to the present invention, a flea saliva protein includes aprotein produced by recombinant DNA methods, as well as proteinsisolated by other methods disclosed in PCT Patent Publication No. WO96/11271.

[0158] Preferred general allergens include those derived from grass,Meadow Fescue, curly dock, plantain, Mexican firebush, lamb's quarters,pigweed, ragweed, sage, elm, cocklebur, box elder, walnut, cottonwood,ash, birch, cedar, oak, mulberry, cockroach, Dermataphagoides,Altemaria, Aspergillus, Cladosporium, Fusarium, Helminthosporium, Mucor,Penicillium, Pullularia, Rhizopus and/or Tricophyton. More preferredgeneral allergens include those derived from Johnson grass, Kentuckyblue grass, meadow fescue, orchard grass, perennial rye grass, red topgrass, timothy grass, Bermuda grass, brome grass, curly dock, Englishplantain, Mexican firebush, lamb's quarters, rough pigweed shortragweed, wormwood sage, American elm, common cocklebur, box elder, blackwalnut, eastern cottonwood, green ash, river birch, red cedar, red oak,red mulberry, cockroach, Dernataphagoides farinae, Altemaria alternata,Aspergillus fumigatus, Cladosporium herbarum, Fusarium vasinfectum,Helminthosporium sativum, Mucor recemosus, Penicillium notatum,Pullularia pullulans, Rhizopus nigricans and/or Tricophyton spp.Preferred parasite antigens include, but are not limited to, helminthantigens, in particular heartworm antigens, such as Di33 (described inU.S. patent application Ser. No. 08/715,628, filed Sep. 18, 1996, byGrieve et al., which is incorporated by reference herein in itsentirety). The term “derived from” refers to a natural allergen of suchplants or organisms (i.e., an allergen directly isolated from suchplants or organisms), as well as, non-natural allergens of such plantsor organisms that posses at least one epitope capable of eliciting animmune response against an allergen (e.g., produced using recombinantDNA technology or by chemical synthesis).

[0159] A feline IgE molecule can be combined with a buffer in which thefeline IgE molecule is solubilized, and/or with a carrier. Suitablebuffers and carriers are known to those skilled in the art. Examples ofsuitable buffers include any buffer in which a feline IgE molecule canfunction to selectively bind to IgE, such as, but not limited to,phosphate buffered saline, water, saline, phosphate buffer, bicarbonatebuffer, HEPES buffer (N-2-hydroxyethylpiperazine-N′-2-ethanesulfonicacid buffered saline), TES buffer (Tris-EDTA buffered saline), Trisbuffer and TAE buffer (Tris-acetate-EDTA). Examples of carriers include,but are not limited to, polymeric matrices, toxoids, and serum albumins,such as bovine serum albumin. Carriers can be mixed with feline IgEmolecules or conjugated (i.e., attached) to feline IgE molecules in sucha manner as to not substantially interfere with the ability of thefeline IgE molecules to selectively bind to Fcε receptor.

[0160] In the methods of the present invention, the compounds hereindescribed in detail can form the active ingredient, and are typicallyadministered in admixture with suitable pharmaceutical diluents,excipients or carriers (collectively referred to herein as “carrier”materials) suitably selected with respect to the intended form ofadministration, that is, oral tablets, capsules, elixirs, syrups and thelike, and consistent with conventional pharmaceutical practices.

[0161] For instance, for oral administration in the form of a tablet orcapsule, the active drug component can be combined with an oral,non-toxic pharmaceutically acceptable inert carrier such as ethanol,glycerol, water, and the like. Moreover, when desired or necessary,suitable binders, lubricants, disintegrating agents and coloring agentscan also be incorporated into the mixture. Suitable binders include,without limitation, starch, gelatin, natural sugars such as glucose orbeta-lactose, corn sweeteners, natural and synthetic gums such asacacia, tragacanth or sodium alginate, carboxymethylcellulose,polyethylene glycol, waxes and the like. Lubricants used in these dosageforms include, without limitation, sodium oleate, sodium stearate,magnesium stearate, sodium benzoate, sodium acetate, sodium chloride,and the like. Disintegrators include, without limitation, starch, methylcellulose, agar, bentonite, xanthan gum and the like.

[0162] For liquid forms the active drug component can be combined insuitably flavored suspending or dispersing agents such as the syntheticand natural gums, for example, tragacanth, acacia, methylcellulose andthe like. Other dispersing agents which may be employed include glycerinand the like. For parenteral administration, sterile suspensions andsolutions are desired. Isotonic preparations which generally containsuitable preservatives are employed when intravenous administration isdesired.

[0163] Topical preparations containing the active drug component can beadmixed with a variety of carrier materials well known in the art, suchas, e.g., alcohols, aloe vera gel, allantoin, glycerine, vitamin A and Eoils, mineral oil, PPG2 myristyl propionate, and the like, to form,e.g., alcoholic solutions, topical cleansers, cleansing creams, skingels, skin lotions, and shampoos in cream or gel formulations. Thecompounds of the present invention can also be administered in the formof liposome delivery systems, such as small unilamellar vesicles, largeunilamellar vesicles and multilamellar vesicles. Liposomes can be formedfrom a variety of phospholipids, such as cholesterol, stearylamine orphosphatidylcholines.

[0164] Compounds of the present invention may also be delivered by theuse of monoclonal antibodies as individual carriers to which thecompound molecules are coupled. The compounds of the present inventionmay also be coupled with soluble polymers as targetable drug carriers.Such polymers can include polyvinyl-pyrrolidone, pyran copolymer,polyhydroxypropylmethacryl-amidephenol,polyhydroxy-ethylaspartamidephenol, or polyethyleneoxidepolylysinesubstituted with palmitoyl residues. Furthermore, the compounds of thepresent invention may be coupled to a class of biodegradable polymersuseful in achieving controlled release of a drug, for example,polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid,polyorthoesters, polyacetals, polydihydro-pyrans, polycyanoacrylates andcross-linked or amphipathic block copolymers of hydrogels.

[0165] In another embodiment of the present invention, there areprovided methods to inhibit or elicit an IgE-mediated immune response,comprising administering a therapeutic compound of the presentinvention.

[0166] One embodiment of the present invention is a method ofimmunotherapy comprising the steps of: (a) administering to an animal aneffective amount of a therapeutic composition of an inhibitor of felineIgE and Fcε receptor complex formation. Suitable therapeuticcompositions and methods of administration methods are disclosed herein.According to the present invention, a therapeutic composition and methodof the present invention can be used to prevent or alleviate symptomsassociated with IgE-mediated biological responses.

[0167] The efficacy of a therapeutic composition of the presentinvention to effect IgE-mediated biological responses can be testedusing standard methods for detecting IgE-mediated immunity including,but not limited to, immediate hypersensitivity, delayedhypersensitivity, antibody-dependent cellular cytotoxicity (ADCC),immune complex activity, mitogenic activity, histamine release assaysand other methods such as those described in Janeway et al., ibid.

[0168] The present invention also provides methods to identify theability of a test compound to interfere with IgE/Fcε receptorinteraction, comprising: contacting the test compound with a protein ofthe present invention; and determining whether the test compound andsaid protein interact.

[0169] In particular, there are provided methods to identify the abilityof a test compound to interfere with IgE/Fcε receptor interactioncomprising: (a) contacting an isolated feline IgE molecule with a testcompound/Fcε receptor containing solution under conditions suitable forformation of an IgE molecule:Fcε receptor complex; and (b) determiningthe ability of the test compound to interfere with IgE/Fcε interactionby detecting the IgE molecule:Fcε receptor complex, the presence of theIgE molecule:Fcε receptor complex indicating the presence of IgE. Apreferred feline IgE molecule is one which a carbohydrate group of thefeline IgE molecule is conjugated to biotin.

[0170] Another embodiment of the present invention is a method toidentify the ability of a test compound to interfere with IgE/Fcεinteraction comprising: (a) contacting a Fcε receptor-bearing cell testcompound and an IgE molecule of the present invention under conditionssuitable for formation of a recombinant cell:IgE complex; and (b)determining the ability of the test compound to interfere with IgE/Fcεreceptor interaction by detecting the recombinant cell:IgE complex, thepresence of the recombinant cell:IgE complex indicating the ability ofthe test compound to interfere with IgE/Fcε receptor interaction.

[0171] A preferred method to detect the ability of the test compound tointerfere with IgE/Fcε receptor interaction comprises: (a) immobilizinga presently-disclosed IgE or a Fcε receptor molecule on a substrate; (b)contacting the IgE or Fcε receptor molecule with the test compound underconditions suitable for formation of an IgE molecule:Fcε receptorcomplex bound to the substrate; (c) removing non-bound material from thesubstrate under conditions that retain Fcε receptor molecule:IgE complexbinding to the substrate; and (d) detecting the presence of the IgEmolecule:Fcε receptor complex.

[0172] Also included are methods to detect IgE, which comprise: (a)immobilizing a test compound on a substrate; (b) contacting the testcompound with a presently-disclosed feline IgE molecule under conditionssuitable for formation of a feline IgE molecule:test compound complexbound to the substrate; (c) removing non-bound material from thesubstrate under conditions that retain feline IgE molecule:test compoundcomplex binding to the substrate; and (d) detecting the presence of thefeline IgE molecule:test compound complex.

[0173] A preferred method to detect IgE comprises: (a) immobilizing Fcεreceptor on a substrate; (b) contacting a test compound with apresently-disclosed feline IgE molecule under conditions suitable forformation of a IgE:test compound complex bound to the Fcε receptor onthe substrate; (c) removing non-bound material from the Fcε receptor onthe substrate under conditions that retain feline IgE molecule:testcompound complex binding to the Fcε receptor on the substrate; and (d)detecting the presence of the feline IgE molecule:test compound complex.

[0174] One embodiment of the present invention is a method to detect IgEnucleic acid which includes the steps of: (a) contacting an isolatedfeline IgE nucleic acid molecule with a putative IgE nucleicacid-containing composition under conditions suitable for formation of afeline IgE nucleic acid molecule:IgE nucleic acid complex; and (b)detecting the presence of IgE nucleic acid by detecting the feline IgEnucleic acid molecule:IgE nucleic acid complex. Presence of such afeline IgE nucleic acid molecule:IgE nucleic acid complex indicates thatthe animal is producing IgE. Preferred IgE to detect using a feline IgEnucleic acid molecule include feline IgE, canine IgE, equine IgE andhuman IgE, with feline IgE being particularly preferred.

[0175] As used herein, canine refers to any member of the dog family,including domestic dogs, wild dogs and zoo dogs. Examples of dogsinclude, but are not limited to, domestic dogs, wild dogs, foxes,wolves, jackals and coyotes. As used herein, equine refers to any memberof the horse family, including horses, donkeys, mules and zebras.

[0176] As used herein, the term “contacting” refers to combining ormixing ingredients, as all of those terms are known in the art.“Formation of a complex” refers to the ability of the molecules to forma stable complex that can be measured (i.e., detected). Binding betweena feline Fcε receptor and a feline IgE molecule is effected underconditions suitable to form a complex; such conditions (e.g.,appropriate concentrations, buffers, temperatures, reaction times) aswell as methods to optimize such conditions are known to those skilledin the art, and examples are disclosed herein. Examples of complexformation conditions are also disclosed in, for example, in Sambrook etal., ibid.

[0177] As used herein, the term “detecting complex formation” refers todetermining if any complex is formed, i.e., assaying for the presence(i.e., existence) of a complex. If complexes are formed, the amount ofcomplexes formed can, but need not be, determined. Complex formation, orselective binding, e.g., between Fcε receptor and feline IgE moleculesin the composition can be measured (i.e., detected, determined) using avariety of methods standard in the art (see, for example, Sambrook etal. ibid.), examples of which are disclosed herein.

[0178] In one embodiment, a test compound of the present method includesa biological sample from an animal. A suitable biological sampleincludes, but is not limited to, a bodily fluid composition or a fewcellular composition. A bodily fluid refers to any fluid that can becollected (i.e., obtained) from an animal, examples of which include,but are not limited to, blood, serum, plasma, urine, tears, aqueoushumor, cerebrospinal fluid (CSF), saliva, lymph, nasal secretions, milkand feces. Such a composition of the present method can, but need notbe, pretreated to remove at least some of the non-IgE isotypes ofimmunoglobulin and/or other proteins, such as albumin, present in thefluid. Such removal can include, but is not limited to, contacting thebodily fluid with a material, such as Protein G, to remove IgGantibodies and/or affinity purifying IgE antibodies from othercomponents of the body fluid by exposing the fluid to, for example,Concanavalin A. In another embodiment, a composition includes collectedbodily fluid that is pretreated to concentrate immunoglobulin containedin the fluid. For example, immunoglobulin contained in a bodily fluidcan be precipitated from other proteins using ammonium sulfate. Apreferred composition of the present method is serum.

[0179] A complex can be detected in a variety of ways including, but notlimited to use of one or more of the following assays: an enzyme-linkedimmunoassay, a radioimmunoassay, a fluorescence immunoassay, achemiluminescent assay, a lateral flow assay, an agglutination assay, aparticulate-based assay (e.g., using particulates such as, but notlimited to, magnetic particles or plastic polymers, such as latex orpolystyrene beads), an immunoprecipitation assay, a BioCore™ assay(e.g., using colloidal gold) and an immunoblotting assay (e.g., awestern blot). Another preferred method is a flow-through assay,examples of which are disclosed in U.S. Pat. No. 4,727,019, issued Feb.23, 1988, by Valkirs et al, which is incorporated by reference in itsentirety. Such assays are well known to those skilled in the art. Assayscan be used to give qualitative or quantitative results depending on howthey are used. Some assays, such as agglutination, particulateseparation, and immunoprecipitation, can be observed visually (e.g.,either by eye or by a machines, such as a densitometer orspectrophotometer) without the need for a detectable marker. In otherassays, conjugation (i.e., attachment) of a detectable marker to thefeline IgE molecule or to a reagent that selectively binds to the felineIgE protein or nucleic acid or to the molecule being detected (describedin more detail below) aids in detecting complex formation. Examples ofdetectable markers include, but are not limited to, a radioactive label,an enzyme, a fluorescent label, a chemiluminescent label, a chromophoriclabel or a ligand. A ligand refers to a molecule that binds selectivelyto another molecule. Preferred detectable markers include, but are notlimited to, fluorescein, a radioisotope, a phosphatase (e.g., alkalinephosphatase), biotin, avidin, a peroxidase (e.g., horseradishperoxidase) and biotin-related compounds or avidin-related compounds(e.g., streptavidin or ImmunoPure® NeutrAvidin available from Pierce,Rockford, Ill.). According to the present invention, a detectable markercan be connected to a feline IgE molecule using, for example, chemicalconjugation or recombinant DNA technology (e.g., connection of a fusionsegment such as that described herein for a metal binding domain; animmunoglobulin binding; a sugar binding domain; and a “tag” domain).Preferably a carbohydrate group of the feline IgE molecule is chemicallyconjugated to biotin.

[0180] In one embodiment, a complex is detected by contacting a testcompound with a feline IgE that is conjugated to a detectable marker. Asuitable detectable marker to conjugate to a feline IgE moleculeincludes, but is not limited to, a radioactive label, a fluorescentlabel, an enzyme label, a chemiluminescent label, a chromophoric labelor a ligand. A detectable marker is conjugated to a feline IgE moleculein such a manner as not to block the ability of the feline IgE moleculeto bind to the compound being detected. Preferably, a feline IgEmolecule is conjugated to biotin.

[0181] In one preferred embodiment, a feline IgE molecule:test compoundcomplex is detected by contacting the complex with an indicator moleculethat selectively binds to a feline IgE molecule of the presentinvention. Examples of such indicator molecule includes, but are notlimited to, an antibody that selectively binds to a feline IgE molecule(referred to herein as an anti-feline IgE antibody) or a compound thatselectively binds to a detectable marker conjugated to a feline IgEmolecule, such as human Fcε receptor, Feline Fcε receptor, or an antigenthat binds to an IgE. A feline IgE molecule conjugated to biotin ispreferably detected using streptavidin, more preferably usingImmunoPure® NeutrAvidin (available from Pierce, Rockford, Ill.).

[0182] In another preferred embodiment, a feline IgE molecule:testcompound complex is detected by contacting the complex with indicatormolecule that selectively binds to an anti-test compound antibody. Asused herein, an anti-test compound antibody includes not only a completeantibody but also any subunit or portion thereof that is capable ofselectively binding to test compound. For example, an anti-test compoundantibody can include an Fab fragment or a F(ab′)₂ fragment, both ofwhich are described in detail in Janeway et al., in Immunobiology, theImmune System in Health and Disease, Garland Publishing, Inc., N.Y.,1996 (which is incorporated herein by this reference in its entirety).

[0183] In one embodiment a complex can be formed and detected insolution. In another embodiment, a complex can be formed in which one ormore members of the complex are immobilized on (e.g., coated onto) asubstrate. Immobilization techniques are known to those skilled in theart. Suitable substrate materials include, but are not limited to,plastic, glass, gel, celluloid, paper, PVDF (poly-vinylidene-fluoride),nylon, nitrocellulose, and particulate materials such as latex,polystyrene, nylon, nitrocellulose, agarose and magnetic resin. Suitableshapes for substrate material include, but are not limited to, a well(e.g., microtiter dish well), a plate, a dipstick, a bead, a lateralflow apparatus, a membrane, a filter, a tube, a dish, a celluloid-typematrix, a magnetic particle, and other particulates. A particularlypreferred substrate comprises an ELISA plate, a dipstick, aradioimmunoassay plate, agarose beads, plastic beads, latex beads,immunoblot membranes and immunoblot papers. In one embodiment, asubstrate, such as a particulate, can include a detectable marker.Another preferred method is a flow-through assay, examples of which aredisclosed in U.S. Pat. No. 4,727,019, issued Feb. 23, 1988, by Valkirset al, which is incorporated by reference in its entirety.

[0184] A preferred method to detect feline IgE molecules of the presentinvention is an immunosorbent assay. An immunoabsorbent assay of thepresent invention comprises a capture molecule and an indicatormolecule. A capture molecule of the present invention binds to an IgE insuch a manner that the IgE is immobilized to a substrate. As such, acapture molecule is preferably immobilized to a substrate of the presentinvention prior to exposure of the capture molecule to a putativeIgE-containing composition. An indicator molecule of the presentinvention detects the presence of an IgE bound to a capture molecule. Assuch, an indicator molecule preferably is not immobilized to the samesubstrate as a capture molecule prior to exposure of the capturemolecule to a putative IgE-containing composition.

[0185] A preferred immunoabsorbent assay method includes a step ofeither: (a) immobilizing a feline IgE molecule on a substrate prior tocontacting a feline IgE molecule with a test compound to form a felineIgE molecule-immobilized substrate; and (b) binding a test compound on asubstrate prior to contacting a feline IgE molecule with a test compoundto form a test compound-bound substrate. Preferably, the substrateincludes a non-coated substrate, a feline IgE molecule-immobilizedsubstrate, an antigen-immobilized substrate or an anti-IgEantibody-immobilized substrate.

[0186] Both a capture molecule and an indicator molecule of the presentinvention are capable of binding to an IgE. Preferably, a capturemolecule binds to a different region of an IgE than an indicatormolecule, thereby allowing a capture molecule to be bound to an IgE atthe same time as an indicator molecule. The use of a reagent as acapture molecule or an indicator molecule depends upon whether themolecule is immobilized to a substrate when the molecule is exposed toan IgE. For example, a feline IgE molecule of the present invention isused as a capture molecule when the feline IgE molecule is bound on asubstrate. Alternatively, a feline IgE molecule is used as an indicatormolecule when the feline IgE molecule is not bound on a substrate.Suitable molecules for use as capture molecules or indicator moleculesinclude, but are not limited to, a feline IgE molecule of the presentinvention, an antigen reagent or an anti-IgE antibody reagent of thepresent invention.

[0187] An immunoabsorbent assay of the present invention can furthercomprise one or more layers and/or types of secondary molecules or otherbinding molecules capable of detecting the presence of an indicatormolecule. For example, an untagged (i.e., not conjugated to a detectablemarker) secondary antibody that selectively binds to an indicatormolecule can be bound to a tagged (i.e., conjugated to a detectablemarker) tertiary antibody that selectively binds to the secondaryantibody. Suitable secondary antibodies, tertiary antibodies and othersecondary or tertiary molecules can be selected by those of skill in theart. Preferred secondary molecules of the present invention include anantigen, an anti-IgE idiotypic antibody and an anti-IgE isotypicantibody. Preferred tertiary molecules can be selected by a skilledartisan based upon the characteristics of the secondary molecule. Thesame strategy is applied for subsequent layers.

[0188] In one embodiment, a feline IgE molecule is used as a capturemolecule by being immobilized on a substrate, such as a microtiter dishwell or a dipstick. A biological sample collected from an animal isapplied to the substrate and incubated under conditions suitable toallow for feline IgE molecule:test compound complex formation bound tothe substrate. Excess non-bound material, if any, is removed from thesubstrate under conditions that retain feline IgE molecule:test compoundcomplex binding to the substrate. An indicator molecule that canselectively bind to a test compound bound to the feline IgE molecule isadded to the substrate and incubated to allow formation of a complexbetween the indicator molecule and the feline IgE molecule:test compoundcomplex. Preferably, the indicator molecule is conjugated to adetectable marker (preferably to an enzyme label, to a colorimetriclabel, to a fluorescent label, to a radioisotope, or to a ligand such asof the biotin or avidin family). Excess indicator molecule is removed, adeveloping agent is added if required, and the substrate is submitted toa detection device for analysis. Preferred test compounds to detect areFc68 R from any animal, antigens or anti-IgE antibodies.

[0189] In one embodiment, an immunosorbent assay of the presentinvention does not utilize a capture molecule. In this embodiment, atest sample is applied to a substrate, such as a microtiter dish well ora dipstick, and incubated under conditions suitable to allow for thetest compound binding to the substrate. Any test compound is immobilizedon the substrate. Excess non-bound material, if any, is removed from thesubstrate under conditions that retain test compound binding to thesubstrate. A feline IgE molecule is added to the substrate and incubatedto allow formation of a complex between the feline IgE molecule and thetest compound. Preferably, the feline IgE molecule is conjugated to adetectable marker (preferably to biotin, an enzyme label or afluorescent label). Excess feline IgE molecule is removed, a developingagent is added if required, and the substrate is submitted to adetection device for analysis. Preferred test compounds to detect areFcε receptor from any animal, antigens or antiIgE antibodies.

[0190] Another preferred method to detect a test compound is a lateralflow assay, examples of which are disclosed in U.S. Pat. No. 5,424,193,issued Jun. 13, 1995, by Pronovost et al.; U.S. Pat. No. 5,415,994,issued May 16, 1995, by Imrich et al; WO 94/29696, published Dec. 22,1994, by Miller et al.; and WO 94/01775, published Jan. 20, 1994, byPawlak et al.; each of these patent publications is incorporated byreference herein in its entirety. In one embodiment, a biological sampleis placed in a lateral flow apparatus that includes the followingcomponents: (a) a support structure defining a flow path; (b) a labelingreagent comprising a feline IgE, the labeling reagent being impregnatedwithin the support structure in a labeling zone; and (c) a capturereagent comprising an anti-feline IgE antibody. The capture reagent islocated downstream of the labeling reagent within a capture zone fluidlyconnected to the labeling zone in such a manner that the labelingreagent can flow from the labeling zone into the capture zone. Thesupport structure comprises a material that does not impede the flow ofthe beads from the labeling zone to the capture zone. Suitable materialsfor use as a support structure include ionic (i.e., anionic or cationic)material. Examples of such a material include, but are not limited to,nitrocellulose (NC), PVDF, carboxymethylcellulose (CM). The supportstructure defines a flow path that is lateral and is divided into zones,namely a labeling zone and a capture zone. The apparatus can furthercomprise a sample receiving zone located along the flow path, morepreferably upstream of the labeling reagent. The flow path in thesupport structure is created by contacting a portion of the supportstructure downstream of the capture zone, preferably at the end of theflow path, to an absorbent capable of absorbing excess liquid from thelabeling and capture zones.

[0191] In this embodiment, the biological sample is applied to thesample receiving zone which includes a portion of the support structure.The labeling zone receives the sample from the sample receiving zonewhich is directed downstream by the flow path. The labeling zonecomprises the feline IgE. A preferred labeling reagent is feline IgEconjugated, either directly or through a linker, to a plastic beadsubstrate, such as to a latex bead. The substrate also includes adetectable marker, preferably a calorimetric marker. Typically, thelabeling reagent is impregnated to the support structure by drying orlyophilization. The sample structure also comprises a capture zonedownstream of the labeling zone. The capture zone receives labelingreagent from the labeling zone which is directed downstream by the flowpath. The capture zone contains the capture reagent, in this case ananti-feline IgE antibody, as disclosed above, that immobilizes the IgEcomplexed to the anti-IgE in the capture zone. The capture reagent ispreferably fixed to the support structure by drying or lyophilizing. Thelabeling reagent accumulates in the capture zone and the accumulation isassessed visually or by an optical detection device.

[0192] Another preferred method is a flow-through assay, examples ofwhich are disclosed in U.S. Pat. No. 4,727,019, issued Feb. 23, 1988, byValkirs et al, which is incorporated by reference in its entirety.

[0193] Also provided by the present invention are methods for inhibitingan immune response to feline IgE, comprising administering a therapeuticcomposition of the present invention in such a manner so as to reducefeline IgE-mediated immune response.

[0194] In another embodiment of the present invention, there areprovided methods for eliciting an immune response to feline IgE,comprising administering an immunogen derived from the feline IgE, or aportion thereof. In particular, a method as above, wherein the portionof the IgE molecule is the constant region is preferred. More preferredis a method as above, wherein the portion of the IgE molecule is the Fcεreceptor-binding region. Most preferred is a method as above wherein theportion of the IgE molecule is SEQ ID NO 2, SEQ ID NO 4 or SEQ ID NO 6.

[0195] The therapeutic compounds and/or compositions can be administeredand formulated as described herein.

[0196] Also included in the present invention are kits comprising thenucleic acids, proteins or inhibitors of the present invention. In broadterms, a kit may contain feline IgE DNA, antibodies to feline IgE, orfeline IgE protein. A kit may be used to detect DNA which hybridizes tofeline IgE DNA or to detect the presence of feline IgE protein orpeptide fragments in a sample. Such characterization is useful for avariety of purposes including but not limited to forensic analyses andepidemiological studies. Alternatively, a kit may contain DNA molecules,RNA molecules, recombinant protein and antibodies of the presentinvention for the purpose of screening and measuring levels of felineIgE DNA, feline IgE RNA or feline IgE protein. The recombinant proteins,DNA molecules, RNA molecules and antibodies lend themselves to theformulation of kits suitable for the detection and typing of feline IgE.All of these kits would comprise a compartmentalized carrier suitable tohold in close confinement at least one container. The carrier may alsofurther comprise reagents such as recombinant feline IgE protein oranti-feline IgE antibodies suitable for detecting feline IgE. Thecarrier may also contain a means for detection such as labeled antigenor enzyme substrates or the like. A preferred kit of the presentinvention further comprises a detection means including one or moreantigens disclosed herein, an antibody capable of selectively binding toan IgE disclosed herein and/or a compound capable of binding to adetectable marker conjugated to a feline IgE protein (e.g., avidin,streptavidin and ImmunoPure® NeutrAvidin when the detectable marker isbiotin). Such antigens preferably induce IgE antibody production inanimals including canines, felines and/or equines.

[0197] In particular, a method and kit of the present invention areuseful for diagnosing abnormal conditions in animals that are associatedwith changing levels of FEc receptor. Particularly preferred conditionsto diagnose include allergies, parasitic infections and neoplasia. Forexample, a method and kit of the present invention are particularlyuseful for detecting flea allergy dermatitis (FAD), when such method orkit includes the use of a flea saliva antigen. FAD is defined as ahypersensitive response to fleabites. Preferably, a putativeIgE-containing composition is obtained from an animal suspected ofhaving FAD. Preferred animals include those disclosed herein, with dogsand cats being more preferred. In addition, methods and kits of thepresent invention are particularly useful for detecting helminthinfection, in particular heartworm infection, when such methods or kitsinclude the use of a helminth antigen, such as Di33. Preferably, aputative IgE-containing composition is obtained from an animal suspectedof having a helminth infection. Preferred animals include thosedisclosed herein, with dogs and cats being more preferred.

[0198] The following examples illustrate the present invention without,however, limiting it. It is to be noted that the Examples include anumber of molecular biology, microbiology, immunology and biochemistrytechniques considered to be known to those skilled in the art.Disclosure of such techniques can be found, for example, in Sambrook etal., ibid., and related references.

EXAMPLE 1 Isolation of a Nucleic Acid Molecule Encoding a Feline IgEKappa Light Chain

[0199] This example describes the isolation, by DNA hybridization, of anucleic acid molecule encoding an IgE kappa light chain subunit fromFelis catus. This nucleic acid molecule was isolated from a felinespleen cDNA library by its ability to hybridize with a ³²P-labelled cDNAencoding the canine IgE kappa light chain subunit.

[0200] A feline spleen cDNA library was prepared as follows. Total RNAwas extracted from spleen material of a cat using anacid-guanidinium-phenol-chloroform method similar to that described byChomzynski, et al, 1987, Anal.Biochem. 162, 156-159. Poly A⁺ selectedRNA was separated from the total RNA population by oligo-dT cellulosechromatography using the mRNA Purification Kit (available from PharmaciaBiotech, Newark, N.J.), according to the method recommended by themanufacturer. A feline spleen library was constructed in lambda-Uni-ZAP™XR vector (available from Stratagene, La Jolla, Calif.) usingStratagene's ZAP-cDNA Synthesis Kit protocol. Approximately 5 μg of polyA+RNA was used to produce the spleen library.

[0201] Using a modification of the protocol described in the cDNASynthesis Kit, the spleen library was screened, using duplicate plaquelifts, with a ³²P-labelled cDNA encoding the canine IgE kappa lightchain subunit. Approximately a million plaques were screened under thefollowing conditions. Filters containing plaques were denatured indenaturation buffer consisting of 0.5 N NaOH and 1.5 M NaCl, andneutralized in neutralization buffer consisting of 1.5 M NaCl and 0.5 MTris-HCl, pH 8.0. Following neutralization, the filters were rinsedbriefly in 2×SSC and subjected to UV crosslinking using, for example, aStratagene UV Stratalinker 1800. The filters were then blocked inhybridization buffer containing 5×SSC, 5×Denhardt's solution, 0.5% SDS,and 100 mg/ml single-stranded DNA for 3 hours at 52° C. The labelledcDNA probe encoding the canine IgE kappa light chain subunit was added,and hybridization was carried out overnight at 52°C. The filters werethen washed in 2×SSC, 0.5% SDS at room temperature for 15 minutes,followed by two washes, for 10 minutes each, in 0.2×SSC, 0.1% SDS, at55° C. The filters were rinsed in 2×SSC, air dried and subjected toautoradiography.

[0202] A plaque purified clone of the feline nucleic acid moleculeencoding the IgE kappa light chain subunit was converted into adouble-stranded DNA molecule using the ExAssist™ helper phage and SOLR™E. coliaccording to the in vivo excision protocol described in theZAP-cDNA Synthesis Kit (available from Stratagene). Double-strandedplasmid DNA was prepared using the Quantum Prep Plasmid Midiprep Kit(available from Bio-Rad, Hercules, Calif.), according to themanufacturer's protocol.

[0203] The plasmid containing the cDNA encoding the feline IgE kappalight chain was sequenced by the Sanger dideoxy chain terminationmethod, using the PRISM™ Ready Dye Terminator Cycle Sequencing Kit withAmpliTaq DNA Polymerase, FS (available from Perkin Elmer Corporation,Norwalk, Conn.). PCR extensions were done in the GeneAmp™ Gel FiltrationCartridge (available from Advanced Genetic Technologies, Gaithersburg,Md.) following their standard protocol. Samples were resuspendedaccording to ABI protocols and were run on a Perkin-Elmer ABI-PRISM™ 377Automated DNA Sequencer. DNA sequence analysis, including thecompilation of sequences and the determination of open reading frames,were performed using the MacVector™ program (available from IBI, NewHaven, Conn.). Protein sequence analysis, including the determination ofmolecular weight and isoelectric point (pI) was performed using the GCG™program (available from Genetics Computer Group, Madison, Wis.).

[0204] Sequence analysis indicated that the nucleic acid moleculeencoding feline IgE kappa light chain was about 954 nucleotides inlength, and, as such, the nucleic acid molecule is referred to herein asnfIgEKLC₉₅₄, Nucleic acid molecule nfIgEKLC₉₅₄ has a coding strand witha nucleic acid sequence of SEQ ID NO: 19 and a complementary strand witha nucleic acid sequence of SEQ ID NO:21. Translation of SEQ ID NO:19indicates that nfIgEKLC₉₅₄ apparently includes a full-length codingregion, with the apparent start and stop codons spanning nucleotides 7through 9 and 733 through 735, respectively, of SEQ ID NO:19. Putativepolyadenylation signals (5′ AATAAA 3′) are located in a region spanningnucleotides 905 through 910 and nucleotides 909 through 914 of SEQ IDNO:19. Translation of SEQ ID NO:19 further indicates that nfIgEKLC₉₅₄encodes a protein of about 242 amino acids, referred to herein asPfIgEKLC₂₄₂, the amino acid sequence of which is presented in SEQ IDNO:20. PfIgEKLC₂₄₂ is encoded by nucleic acid molecule nfIgEKLC₇₂₆,which consists of a coding strand having SEQ ID NO:23 and acomplementary strand having SEQ ID NO:24. SEQ ID NO:20 predicts afull-length feline IgE kappa light chain protein with an estimatedmolecular weight of about 26.7 kD and an estimated pI of about 6.5.Analysis of SEQ ID NO:20 suggests the presence of a signal peptideencoded by a stretch of amino acids spanning from about amino acid 1through amino acid 20. The proposed mature protein, referred to hereinas PfIgEKLC₂₂₂, contains about 222 amino acids which is representedherein as SEQ ID NO:26, encoded by nucleic acid molecule nfIgEKLC₆₆₆,consisting of a coding strand with a nucleic acid sequence of SEQ IDNO:25 and a complementary strand with a nucleic acid sequence of SEQ IDNO:27. SEQ ID NO:26 predicts a mature feline IgE kappa light chainprotein with an estimated molecular weight of about 24.5 kD and anestimated pI of about 6.13.

[0205] A BLASTp search was performed by searching the NIH database athttp://www.ncbi.nlm.nih.gov/BLAST/. The protein search was performedusing SEQ ID NO:20, which showed significant homology to several kappalight chain proteins. The highest scoring match of the homology searchat the amino acid level was Rattus norvegicus (Norway rat) Ig kappalight chain (AAB03702), which was about 61% identical with SEQ ID NO:20.At the nucleotide level, the search was performed using SEQ ID NO:19,which was most similar (sharing 84% nucleotide sequence identity) to thecat kappa light chain sequences (M90809).

EXAMPLE 2 Isolation of a Nucleic Acid Molecule Encoding a Feline IgEEpsilon Heavy Chain

[0206] This example describes the isolation, by DNA hybridization, of anucleic acid molecule encoding an IgE epsilon heavy chain subunit fromFelis catus. This nucleic acid molecule was isolated from a felinespleen cDNA library in a manner similar to that described in Example 1except that the labelled probe used was a ³²P-labelled cDNA encoding thecanine IgE epsilon heavy chain subunit.

[0207] A plaque purified clone of the feline nucleic acid moleculeencoding the IgE epsilon heavy chain subunit was converted into adouble-stranded plasmid as described in Example 1 and submitted tosequence analysis as described in Example 1.

[0208] Sequence analysis indicated that the isolated nucleic acidmolecule encoding feline IgE epsilon heavy chain was about 1613nucleotides, and, as such, the nucleic acid molecule is referred toherein as nfIgEEHC₁₆₁₃. Nucleic acid molecule nfIgEEHC₁₆₁₃ has a codingstrand with a nucleic acid sequence of SEQ ID NO:1 and a complementarystrand with a nucleic acid sequence of SEQ ID NO:3. Translation of SEQID NO:1 indicates that nfIgEEHC₁₆₁₃ apparently includes a partial codingregion, with an apparent stop codon spanning nucleotides 1489 through1491 of SEQ ID NO:1. A putative polyadenylation signal (5′ AATAAA 3′) islocated in a region spanning nucleotides 1569 through 1574 of SEQ IDNO:1. Translation of SEQ ID NO:1 further indicates that nfIgEEHC₁₆₁₃encodes a protein of about 496 amino acids, referred to herein asPfIgEEHC₄₉₆, the amino acid sequence of which is presented in SEQ IDNO:2. PfIgEEHC₄₉₆ is encoded by nucleic acid molecule nfIgEEHC_(1488,)which consists of a coding strand having SEQ ID NO:31 (with theexception of an g instead of an a at base 205 of SEQ ID NO 31) and acomplementary strand having SEQ ID NO:32 (with an exception of a cinstead of a t at the complementary position). SEQ ID NO:2 predicts apartial feline IgE epsilon heavy chain protein with an estimatedmolecular weight of about 54.4 kD and an estimated pIof about 6.84Analysis of SEQ ID NO:2 suggests that the feline IgE epsilon heavy chainprotein includes a partial variable region and an apparent full-lengthconstant region. The amino acid sequence of the partial variable regionis denoted by SEQ ID NO:17 and is encoded by nucleic acid sequence SEQID NO:16, the complement of which is SEQ ID NO:18. The amino acidsequence of the constant region is denoted by SEQ ID NO:14 and isencoded by nucleic acid sequence SEQ ID NO:33 (with noted exceptionabove), the complement of which is SEQ ID NO:34 (with noted exceptionabove). It is to be noted that SEQ ID NO:13 also encodes SEQ ID NO:14,but also contains a 3′ untranslated region.

[0209] A BLASTp search was performed by searching the NIH database athttp://www.ncbi.nlm.nih.gov/BLAST/. The protein search was performedusing SEQ ID NO:2, which showed significant homology to several IgEepsilon heavy chain proteins. The highest scoring match of the homologysearch at the amino acid level was canine IgE epsilon heavy chain(Accession number 598109), which was about 76% identical with SEQ IDNO:2. At the nucleotide level, the search was performed using SEQ IDNO:1, which was most similar to accession number L36872, canine IgEepsilon heavy chain DNA, the percent identity being 82%.

EXAMPLE 3 Antibodies to Feline IgE Heavy and Light Chains

[0210] Feline IgE protein was prepared by passing cat sera through anaffinity column of mouse anti-dog IgE monoclonal antibody (mab) producedby the cell line H-47 (available from Custom Mab, West Sacramento,Calif.) bound to Sepharose 4B. Protein retained on the column was elutedwith 0.1M Glycine-HCl pH 2.8. The eluted protein was diluted 1:5 with 10mM Tris-HCl pH 8.0 and applied to a Q-Sepharose column. The column wasthen eluted sequentially with 0.1M, 0.2M, 0.3M, 0.4M, 0.5M, 0.6M and1.0M of Tris-HCl pH 8.0 and fractions were collected. The 0.2M and 0.3Mfractions were pooled and applied to another affinity column comprisinggoat anti-cat IgG-Sepharose column (available from Kirkgaard & Perry,Gaithersberg, Md.) bound to Sepharose 4B. The flow-through was collectedand found to contain purified cat IgE.

[0211] Two Balb/c mice were immunized in the footpad with 30 μg of thepurified feline IgE suspended in phosphate buffered saline (PBS) andFreund's complete adjuvant. A boost of 30 μg feline IgE was given inPBS/Freund's incomplete adjuvant in the footpad 14 days afterimmunization. Sera was tested for presence of anti-feline IgE antibodies21 days after immunization. The mouse exhibiting the highest titeragainst feline IgE was boosted with 10 μg antigen in PBS, intravenously42 days after immunization. Splenocytes were harvested three days laterand fused with mouse SP2/0 myeloma cells at mid-log growth phase usingPEG.

[0212] Cells were cultured in RPMI media containing 20% fetal bovineserum, 10% thymocyte conditioned media, 2 mM L-glutamine, 1 mM sodiumpyruvate, 60 μM β-mercaptoethanol and hybrids were selected by adding100 μ hypoxanthine, 10 μM thymidine and 0.4 μM aminopterin. Wellscontaining hybridoma colonies were tested for anti-IgE monoclonalantibody production using either feline IgE, IgG and IgM by ELISA usingstandard techniques.

[0213] Eight monoclonal antibodies that bind specifically to feline IgEwere generated and are referred to as H-100, H-101, H-102, H-103, H-106,H-99, H-104, and H-107; H-100, H-101H-102, H-103 and H-106 bind tofeline IgE and do not bind to IgG or IgM. These five antibodies also donot react with canine or human IgE. In addition, three monoclonalantibodies reactive to feline IgE light chain were produced and arereferred to as H-99, H-104, H-107.

1 34 1 1613 DNA Felis catus CDS (1)..(1491) 1 gca tat att agt agt ggaggt aac aca gac tac gca gac tcc gtg aag 48 Ala Tyr Ile Ser Ser Gly GlyAsn Thr Asp Tyr Ala Asp Ser Val Lys 1 5 10 15 ggc cga ttc tcc atc tccaga gac aac gcc aag aac acg ctg tat ctg 96 Gly Arg Phe Ser Ile Ser ArgAsp Asn Ala Lys Asn Thr Leu Tyr Leu 20 25 30 cag atg acc agc ctc aag accgag gac acg gcc aca tat tac tgt gca 144 Gln Met Thr Ser Leu Lys Thr GluAsp Thr Ala Thr Tyr Tyr Cys Ala 35 40 45 aga ggg act ggt gta ata ccg gactac tgg ggc cag gga gcc ctg gtg 192 Arg Gly Thr Gly Val Ile Pro Asp TyrTrp Gly Gln Gly Ala Leu Val 50 55 60 acg gtg tcc tca gcc tcc atc cag gccccc ctc gtc ttc ccc ttg gcc 240 Thr Val Ser Ser Ala Ser Ile Gln Ala ProLeu Val Phe Pro Leu Ala 65 70 75 80 acc tgc tgc aaa ggc acc atc gcc actgcc ccg tcc gtg aca ctg ggc 288 Thr Cys Cys Lys Gly Thr Ile Ala Thr AlaPro Ser Val Thr Leu Gly 85 90 95 tgc ctg gtc acg ggc tac ttc ccg atg ccggtg act gtg acc tgg gat 336 Cys Leu Val Thr Gly Tyr Phe Pro Met Pro ValThr Val Thr Trp Asp 100 105 110 gca agg tcc ctg aac aag agc gtc gtg accctc ccc gcc acc ctc cag 384 Ala Arg Ser Leu Asn Lys Ser Val Val Thr LeuPro Ala Thr Leu Gln 115 120 125 gag acc tct ggc ctc tac acc acc acc agccac gtg acc gtc tcg ggc 432 Glu Thr Ser Gly Leu Tyr Thr Thr Thr Ser HisVal Thr Val Ser Gly 130 135 140 gag tgg gcc aaa cag aag ttc acc tgc agtgtg gct cac gcg gag tcc 480 Glu Trp Ala Lys Gln Lys Phe Thr Cys Ser ValAla His Ala Glu Ser 145 150 155 160 ccc acc atc aac aag acc gtc agt gcgtgt acc atg aac ttc att ccc 528 Pro Thr Ile Asn Lys Thr Val Ser Ala CysThr Met Asn Phe Ile Pro 165 170 175 ccc acc gtg aag ctc ttc cac tcc tcctgt aac ccc ctc ggt gac acc 576 Pro Thr Val Lys Leu Phe His Ser Ser CysAsn Pro Leu Gly Asp Thr 180 185 190 ggt agc acc atc cag ctc ctg tgc ctcatc tcc ggc tac gtc cca ggt 624 Gly Ser Thr Ile Gln Leu Leu Cys Leu IleSer Gly Tyr Val Pro Gly 195 200 205 gac atg gag gtc acc tgg ctg gtg gatggg cag aag gcc acg aac ata 672 Asp Met Glu Val Thr Trp Leu Val Asp GlyGln Lys Ala Thr Asn Ile 210 215 220 ttc cca tac act gcc ccc ggc aag caggag ggc aag gtg acc tcc acc 720 Phe Pro Tyr Thr Ala Pro Gly Lys Gln GluGly Lys Val Thr Ser Thr 225 230 235 240 cac agc gag ctc aac atc acg cagggt gag tgg gtg tcc caa aag acc 768 His Ser Glu Leu Asn Ile Thr Gln GlyGlu Trp Val Ser Gln Lys Thr 245 250 255 tac act tgc cag gtc acc tat caaggc ttc acc ttt gag gac cac gct 816 Tyr Thr Cys Gln Val Thr Tyr Gln GlyPhe Thr Phe Glu Asp His Ala 260 265 270 cgc aag tgc aca gag tct gac ccccga ggt gtg agc acc tac ttg agc 864 Arg Lys Cys Thr Glu Ser Asp Pro ArgGly Val Ser Thr Tyr Leu Ser 275 280 285 ccg ccc agc cct ctt gac ctg tacgtc cac aag tcg ccc aag atc acc 912 Pro Pro Ser Pro Leu Asp Leu Tyr ValHis Lys Ser Pro Lys Ile Thr 290 295 300 tgc ctg gtg gtg gac ctg gcc aacaca gac ggc atg atc ctg acc tgg 960 Cys Leu Val Val Asp Leu Ala Asn ThrAsp Gly Met Ile Leu Thr Trp 305 310 315 320 tcg cgg gag aat ggg gag tctgtg cac cca gac ccg atg gtc aag aag 1008 Ser Arg Glu Asn Gly Glu Ser ValHis Pro Asp Pro Met Val Lys Lys 325 330 335 act cag tac aac ggg aca atcacc gtc act tcc acc ctg cct gtg gat 1056 Thr Gln Tyr Asn Gly Thr Ile ThrVal Thr Ser Thr Leu Pro Val Asp 340 345 350 gcc act gac tgg gtt gag ggggag acc tac caa tgc aag gtg acc cat 1104 Ala Thr Asp Trp Val Glu Gly GluThr Tyr Gln Cys Lys Val Thr His 355 360 365 cca gac ctg ccc aag gac atcgtg cgc tcc att gcc aaa gcc ccc ggc 1152 Pro Asp Leu Pro Lys Asp Ile ValArg Ser Ile Ala Lys Ala Pro Gly 370 375 380 cgg cgt ttc ccc ccg gag gtgtac gtg ttc ctg ccg ccc gag ggg gag 1200 Arg Arg Phe Pro Pro Glu Val TyrVal Phe Leu Pro Pro Glu Gly Glu 385 390 395 400 ccg aag acc aag gac aaagtc att ctc acg tgc ctg atc cag aac ttc 1248 Pro Lys Thr Lys Asp Lys ValIle Leu Thr Cys Leu Ile Gln Asn Phe 405 410 415 ttt ccc ccg gac atc tcggtg caa tgg ctt cac aac gac agc cct gtt 1296 Phe Pro Pro Asp Ile Ser ValGln Trp Leu His Asn Asp Ser Pro Val 420 425 430 cgg aca gaa cag cag gccacc acg tgg ccc cac aag gcc acc ggc ccc 1344 Arg Thr Glu Gln Gln Ala ThrThr Trp Pro His Lys Ala Thr Gly Pro 435 440 445 agc cca gcc ttc ttt gtcttc agt cgc ctt gag gtc agc cgg gca gac 1392 Ser Pro Ala Phe Phe Val PheSer Arg Leu Glu Val Ser Arg Ala Asp 450 455 460 tgg gag cag agg gat gtgttc acc tgc caa gtg gtg cac gag gcg ctg 1440 Trp Glu Gln Arg Asp Val PheThr Cys Gln Val Val His Glu Ala Leu 465 470 475 480 cct ggc ttt agg acgctc aag aaa tcc gtg tcc aaa aac cct ggt aaa 1488 Pro Gly Phe Arg Thr LeuLys Lys Ser Val Ser Lys Asn Pro Gly Lys 485 490 495 tga tgcccacccctccccccaga gctccatcct gctggggcgg gggaggggcc 1541 ggccggacct gccggtctgttgttgtcaat aaacactgca gtgcctgcct cagaaaaaaa 1601 aaaaaaaaaa aa 1613 2496 PRT Felis catus 2 Ala Tyr Ile Ser Ser Gly Gly Asn Thr Asp Tyr AlaAsp Ser Val Lys 1 5 10 15 Gly Arg Phe Ser Ile Ser Arg Asp Asn Ala LysAsn Thr Leu Tyr Leu 20 25 30 Gln Met Thr Ser Leu Lys Thr Glu Asp Thr AlaThr Tyr Tyr Cys Ala 35 40 45 Arg Gly Thr Gly Val Ile Pro Asp Tyr Trp GlyGln Gly Ala Leu Val 50 55 60 Thr Val Ser Ser Ala Ser Ile Gln Ala Pro LeuVal Phe Pro Leu Ala 65 70 75 80 Thr Cys Cys Lys Gly Thr Ile Ala Thr AlaPro Ser Val Thr Leu Gly 85 90 95 Cys Leu Val Thr Gly Tyr Phe Pro Met ProVal Thr Val Thr Trp Asp 100 105 110 Ala Arg Ser Leu Asn Lys Ser Val ValThr Leu Pro Ala Thr Leu Gln 115 120 125 Glu Thr Ser Gly Leu Tyr Thr ThrThr Ser His Val Thr Val Ser Gly 130 135 140 Glu Trp Ala Lys Gln Lys PheThr Cys Ser Val Ala His Ala Glu Ser 145 150 155 160 Pro Thr Ile Asn LysThr Val Ser Ala Cys Thr Met Asn Phe Ile Pro 165 170 175 Pro Thr Val LysLeu Phe His Ser Ser Cys Asn Pro Leu Gly Asp Thr 180 185 190 Gly Ser ThrIle Gln Leu Leu Cys Leu Ile Ser Gly Tyr Val Pro Gly 195 200 205 Asp MetGlu Val Thr Trp Leu Val Asp Gly Gln Lys Ala Thr Asn Ile 210 215 220 PhePro Tyr Thr Ala Pro Gly Lys Gln Glu Gly Lys Val Thr Ser Thr 225 230 235240 His Ser Glu Leu Asn Ile Thr Gln Gly Glu Trp Val Ser Gln Lys Thr 245250 255 Tyr Thr Cys Gln Val Thr Tyr Gln Gly Phe Thr Phe Glu Asp His Ala260 265 270 Arg Lys Cys Thr Glu Ser Asp Pro Arg Gly Val Ser Thr Tyr LeuSer 275 280 285 Pro Pro Ser Pro Leu Asp Leu Tyr Val His Lys Ser Pro LysIle Thr 290 295 300 Cys Leu Val Val Asp Leu Ala Asn Thr Asp Gly Met IleLeu Thr Trp 305 310 315 320 Ser Arg Glu Asn Gly Glu Ser Val His Pro AspPro Met Val Lys Lys 325 330 335 Thr Gln Tyr Asn Gly Thr Ile Thr Val ThrSer Thr Leu Pro Val Asp 340 345 350 Ala Thr Asp Trp Val Glu Gly Glu ThrTyr Gln Cys Lys Val Thr His 355 360 365 Pro Asp Leu Pro Lys Asp Ile ValArg Ser Ile Ala Lys Ala Pro Gly 370 375 380 Arg Arg Phe Pro Pro Glu ValTyr Val Phe Leu Pro Pro Glu Gly Glu 385 390 395 400 Pro Lys Thr Lys AspLys Val Ile Leu Thr Cys Leu Ile Gln Asn Phe 405 410 415 Phe Pro Pro AspIle Ser Val Gln Trp Leu His Asn Asp Ser Pro Val 420 425 430 Arg Thr GluGln Gln Ala Thr Thr Trp Pro His Lys Ala Thr Gly Pro 435 440 445 Ser ProAla Phe Phe Val Phe Ser Arg Leu Glu Val Ser Arg Ala Asp 450 455 460 TrpGlu Gln Arg Asp Val Phe Thr Cys Gln Val Val His Glu Ala Leu 465 470 475480 Pro Gly Phe Arg Thr Leu Lys Lys Ser Val Ser Lys Asn Pro Gly Lys 485490 495 3 1613 DNA Felis catus 3 tttttttttt tttttttttc tgaggcaggcactgcagtgt ttattgacaa caacagaccg 60 gcaggtccgg ccggcccctc ccccgccccagcaggatgga gctctggggg gaggggtggg 120 catcatttac cagggttttt ggacacggatttcttgagcg tcctaaagcc aggcagcgcc 180 tcgtgcacca cttggcaggt gaacacatccctctgctccc agtctgcccg gctgacctca 240 aggcgactga agacaaagaa ggctgggctggggccggtgg ccttgtgggg ccacgtggtg 300 gcctgctgtt ctgtccgaac agggctgtcgttgtgaagcc attgcaccga gatgtccggg 360 ggaaagaagt tctggatcag gcacgtgagaatgactttgt ccttggtctt cggctccccc 420 tcgggcggca ggaacacgta cacctccggggggaaacgcc ggccgggggc tttggcaatg 480 gagcgcacga tgtccttggg caggtctggatgggtcacct tgcattggta ggtctccccc 540 tcaacccagt cagtggcatc cacaggcagggtggaagtga cggtgattgt cccgttgtac 600 tgagtcttct tgaccatcgg gtctgggtgcacagactccc cattctcccg cgaccaggtc 660 aggatcatgc cgtctgtgtt ggccaggtccaccaccaggc aggtgatctt gggcgacttg 720 tggacgtaca ggtcaagagg gctgggcgggctcaagtagg tgctcacacc tcgggggtca 780 gactctgtgc acttgcgagc gtggtcctcaaaggtgaagc cttgataggt gacctggcaa 840 gtgtaggtct tttgggacac ccactcaccctgcgtgatgt tgagctcgct gtgggtggag 900 gtcaccttgc cctcctgctt gccgggggcagtgtatggga atatgttcgt ggccttctgc 960 ccatccacca gccaggtgac ctccatgtcacctgggacgt agccggagat gaggcacagg 1020 agctggatgg tgctaccggt gtcaccgagggggttacagg aggagtggaa gagcttcacg 1080 gtggggggaa tgaagttcat ggtacacgcactgacggtct tgttgatggt gggggactcc 1140 gcgtgagcca cactgcaggt gaacttctgtttggcccact cgcccgagac ggtcacgtgg 1200 ctggtggtgg tgtagaggcc agaggtctcctggagggtgg cggggagggt cacgacgctc 1260 ttgttcaggg accttgcatc ccaggtcacagtcaccggca tcgggaagta gcccgtgacc 1320 aggcagccca gtgtcacgga cggggcagtggcgatggtgc ctttgcagca ggtggccaag 1380 gggaagacga ggggggcctg gatggaggctgaggacaccg tcaccagggc tccctggccc 1440 cagtagtccg gtattacacc agtccctcttgcacagtaat atgtggccgt gtcctcggtc 1500 ttgaggctgg tcatctgcag atacagcgtgttcttggcgt tgtctctgga gatggagaat 1560 cggcccttca cggagtctgc gtagtctgtgttacctccac tactaatata tgc 1613 4 36 DNA Felis catus CDS (1)..(36) 4 agccct ctt gac ctg tac gtc cac aag tcg ccc aag 36 Ser Pro Leu Asp Leu TyrVal His Lys Ser Pro Lys 1 5 10 5 12 PRT Felis catus 5 Ser Pro Leu AspLeu Tyr Val His Lys Ser Pro Lys 1 5 10 6 36 DNA Felis catus 6 cttgggcgacttgtggacgt acaggtcaag agggct 36 7 54 DNA Felis catus CDS (1)..(54) 7 agcccg ccc agc cct ctt gac ctg tac gtc cac aag tcg ccc aag atc 48 Ser ProPro Ser Pro Leu Asp Leu Tyr Val His Lys Ser Pro Lys Ile 1 5 10 15 acctgc 54 Thr Cys 8 18 PRT Felis catus 8 Ser Pro Pro Ser Pro Leu Asp LeuTyr Val His Lys Ser Pro Lys Ile 1 5 10 15 Thr Cys 9 54 DNA Felis catus 9gcaggtgatc ttgggcgact tgtggacgta caggtcaaga gggctgggcg ggct 54 10 78 DNAFelis catus CDS (1)..(78) 10 agc acc tac ttg agc ccg ccc agc cct ctt gacctg tac gtc cac aag 48 Ser Thr Tyr Leu Ser Pro Pro Ser Pro Leu Asp LeuTyr Val His Lys 1 5 10 15 tcg ccc aag atc acc tgc ctg gtg gtg gac 78 SerPro Lys Ile Thr Cys Leu Val Val Asp 20 25 11 26 PRT Felis catus 11 SerThr Tyr Leu Ser Pro Pro Ser Pro Leu Asp Leu Tyr Val His Lys 1 5 10 15Ser Pro Lys Ile Thr Cys Leu Val Val Asp 20 25 12 78 DNA Felis catus 12gtccaccacc aggcaggtga tcttgggcga cttgtggacg tacaggtcaa gagggctggg 60cgggctcaag taggtgct 78 13 1418 DNA Felis catus CDS (1)..(1293) 13 gtgtcc tca gcc tcc atc cag gcc ccc ctc gtc ttc ccc ttg gcc acc 48 Val SerSer Ala Ser Ile Gln Ala Pro Leu Val Phe Pro Leu Ala Thr 1 5 10 15 tgctgc aaa ggc acc atc gcc act gcc ccg tcc gtg aca ctg ggc tgc 96 Cys CysLys Gly Thr Ile Ala Thr Ala Pro Ser Val Thr Leu Gly Cys 20 25 30 ctg gtcacg ggc tac ttc ccg atg ccg gtg act gtg acc tgg gat gca 144 Leu Val ThrGly Tyr Phe Pro Met Pro Val Thr Val Thr Trp Asp Ala 35 40 45 agg tcc ctgaac aag agc gtc gtg acc ctc ccc gcc acc ctc cag gag 192 Arg Ser Leu AsnLys Ser Val Val Thr Leu Pro Ala Thr Leu Gln Glu 50 55 60 acc tct ggc ctctac acc acc acc agc cac gtg acc gtc tcg ggc gag 240 Thr Ser Gly Leu TyrThr Thr Thr Ser His Val Thr Val Ser Gly Glu 65 70 75 80 tgg gcc aaa cagaag ttc acc tgc agt gtg gct cac gcg gag tcc ccc 288 Trp Ala Lys Gln LysPhe Thr Cys Ser Val Ala His Ala Glu Ser Pro 85 90 95 acc atc aac aag accgtc agt gcg tgt acc atg aac ttc att ccc ccc 336 Thr Ile Asn Lys Thr ValSer Ala Cys Thr Met Asn Phe Ile Pro Pro 100 105 110 acc gtg aag ctc ttccac tcc tcc tgt aac ccc ctc ggt gac acc ggt 384 Thr Val Lys Leu Phe HisSer Ser Cys Asn Pro Leu Gly Asp Thr Gly 115 120 125 agc acc atc cag ctcctg tgc ctc atc tcc ggc tac gtc cca ggt gac 432 Ser Thr Ile Gln Leu LeuCys Leu Ile Ser Gly Tyr Val Pro Gly Asp 130 135 140 atg gag gtc acc tggctg gtg gat ggg cag aag gcc acg aac ata ttc 480 Met Glu Val Thr Trp LeuVal Asp Gly Gln Lys Ala Thr Asn Ile Phe 145 150 155 160 cca tac act gccccc ggc aag cag gag ggc aag gtg acc tcc acc cac 528 Pro Tyr Thr Ala ProGly Lys Gln Glu Gly Lys Val Thr Ser Thr His 165 170 175 agc gag ctc aacatc acg cag ggt gag tgg gtg tcc caa aag acc tac 576 Ser Glu Leu Asn IleThr Gln Gly Glu Trp Val Ser Gln Lys Thr Tyr 180 185 190 act tgc cag gtcacc tat caa ggc ttc acc ttt gag gac cac gct cgc 624 Thr Cys Gln Val ThrTyr Gln Gly Phe Thr Phe Glu Asp His Ala Arg 195 200 205 aag tgc aca gagtct gac ccc cga ggt gtg agc acc tac ttg agc ccg 672 Lys Cys Thr Glu SerAsp Pro Arg Gly Val Ser Thr Tyr Leu Ser Pro 210 215 220 ccc agc cct cttgac ctg tac gtc cac aag tcg ccc aag atc acc tgc 720 Pro Ser Pro Leu AspLeu Tyr Val His Lys Ser Pro Lys Ile Thr Cys 225 230 235 240 ctg gtg gtggac ctg gcc aac aca gac ggc atg atc ctg acc tgg tcg 768 Leu Val Val AspLeu Ala Asn Thr Asp Gly Met Ile Leu Thr Trp Ser 245 250 255 cgg gag aatggg gag tct gtg cac cca gac ccg atg gtc aag aag act 816 Arg Glu Asn GlyGlu Ser Val His Pro Asp Pro Met Val Lys Lys Thr 260 265 270 cag tac aacggg aca atc acc gtc act tcc acc ctg cct gtg gat gcc 864 Gln Tyr Asn GlyThr Ile Thr Val Thr Ser Thr Leu Pro Val Asp Ala 275 280 285 act gac tgggtt gag ggg gag acc tac caa tgc aag gtg acc cat cca 912 Thr Asp Trp ValGlu Gly Glu Thr Tyr Gln Cys Lys Val Thr His Pro 290 295 300 gac ctg cccaag gac atc gtg cgc tcc att gcc aaa gcc ccc ggc cgg 960 Asp Leu Pro LysAsp Ile Val Arg Ser Ile Ala Lys Ala Pro Gly Arg 305 310 315 320 cgt ttcccc ccg gag gtg tac gtg ttc ctg ccg ccc gag ggg gag ccg 1008 Arg Phe ProPro Glu Val Tyr Val Phe Leu Pro Pro Glu Gly Glu Pro 325 330 335 aag accaag gac aaa gtc att ctc acg tgc ctg atc cag aac ttc ttt 1056 Lys Thr LysAsp Lys Val Ile Leu Thr Cys Leu Ile Gln Asn Phe Phe 340 345 350 ccc ccggac atc tcg gtg caa tgg ctt cac aac gac agc cct gtt cgg 1104 Pro Pro AspIle Ser Val Gln Trp Leu His Asn Asp Ser Pro Val Arg 355 360 365 aca gaacag cag gcc acc acg tgg ccc cac aag gcc acc ggc ccc agc 1152 Thr Glu GlnGln Ala Thr Thr Trp Pro His Lys Ala Thr Gly Pro Ser 370 375 380 cca gccttc ttt gtc ttc agt cgc ctt gag gtc agc cgg gca gac tgg 1200 Pro Ala PhePhe Val Phe Ser Arg Leu Glu Val Ser Arg Ala Asp Trp 385 390 395 400 gagcag agg gat gtg ttc acc tgc caa gtg gtg cac gag gcg ctg cct 1248 Glu GlnArg Asp Val Phe Thr Cys Gln Val Val His Glu Ala Leu Pro 405 410 415 ggcttt agg acg ctc aag aaa tcc gtg tcc aaa aac cct ggt aaa 1293 Gly Phe ArgThr Leu Lys Lys Ser Val Ser Lys Asn Pro Gly Lys 420 425 430 tgatgcccacccctcccccc agagctccat cctgctgggg cgggggaggg gccggccgga 1353 cctgccggtctgttgttgtc aataaacact gcagtgcctg cctcagaaaa aaaaaaaaaa 1413 aaaaa 141814 431 PRT Felis catus 14 Val Ser Ser Ala Ser Ile Gln Ala Pro Leu ValPhe Pro Leu Ala Thr 1 5 10 15 Cys Cys Lys Gly Thr Ile Ala Thr Ala ProSer Val Thr Leu Gly Cys 20 25 30 Leu Val Thr Gly Tyr Phe Pro Met Pro ValThr Val Thr Trp Asp Ala 35 40 45 Arg Ser Leu Asn Lys Ser Val Val Thr LeuPro Ala Thr Leu Gln Glu 50 55 60 Thr Ser Gly Leu Tyr Thr Thr Thr Ser HisVal Thr Val Ser Gly Glu 65 70 75 80 Trp Ala Lys Gln Lys Phe Thr Cys SerVal Ala His Ala Glu Ser Pro 85 90 95 Thr Ile Asn Lys Thr Val Ser Ala CysThr Met Asn Phe Ile Pro Pro 100 105 110 Thr Val Lys Leu Phe His Ser SerCys Asn Pro Leu Gly Asp Thr Gly 115 120 125 Ser Thr Ile Gln Leu Leu CysLeu Ile Ser Gly Tyr Val Pro Gly Asp 130 135 140 Met Glu Val Thr Trp LeuVal Asp Gly Gln Lys Ala Thr Asn Ile Phe 145 150 155 160 Pro Tyr Thr AlaPro Gly Lys Gln Glu Gly Lys Val Thr Ser Thr His 165 170 175 Ser Glu LeuAsn Ile Thr Gln Gly Glu Trp Val Ser Gln Lys Thr Tyr 180 185 190 Thr CysGln Val Thr Tyr Gln Gly Phe Thr Phe Glu Asp His Ala Arg 195 200 205 LysCys Thr Glu Ser Asp Pro Arg Gly Val Ser Thr Tyr Leu Ser Pro 210 215 220Pro Ser Pro Leu Asp Leu Tyr Val His Lys Ser Pro Lys Ile Thr Cys 225 230235 240 Leu Val Val Asp Leu Ala Asn Thr Asp Gly Met Ile Leu Thr Trp Ser245 250 255 Arg Glu Asn Gly Glu Ser Val His Pro Asp Pro Met Val Lys LysThr 260 265 270 Gln Tyr Asn Gly Thr Ile Thr Val Thr Ser Thr Leu Pro ValAsp Ala 275 280 285 Thr Asp Trp Val Glu Gly Glu Thr Tyr Gln Cys Lys ValThr His Pro 290 295 300 Asp Leu Pro Lys Asp Ile Val Arg Ser Ile Ala LysAla Pro Gly Arg 305 310 315 320 Arg Phe Pro Pro Glu Val Tyr Val Phe LeuPro Pro Glu Gly Glu Pro 325 330 335 Lys Thr Lys Asp Lys Val Ile Leu ThrCys Leu Ile Gln Asn Phe Phe 340 345 350 Pro Pro Asp Ile Ser Val Gln TrpLeu His Asn Asp Ser Pro Val Arg 355 360 365 Thr Glu Gln Gln Ala Thr ThrTrp Pro His Lys Ala Thr Gly Pro Ser 370 375 380 Pro Ala Phe Phe Val PheSer Arg Leu Glu Val Ser Arg Ala Asp Trp 385 390 395 400 Glu Gln Arg AspVal Phe Thr Cys Gln Val Val His Glu Ala Leu Pro 405 410 415 Gly Phe ArgThr Leu Lys Lys Ser Val Ser Lys Asn Pro Gly Lys 420 425 430 15 1418 DNAFelis catus 15 tttttttttt tttttttttc tgaggcaggc actgcagtgt ttattgacaacaacagaccg 60 gcaggtccgg ccggcccctc ccccgcccca gcaggatgga gctctggggggaggggtggg 120 catcatttac cagggttttt ggacacggat ttcttgagcg tcctaaagccaggcagcgcc 180 tcgtgcacca cttggcaggt gaacacatcc ctctgctccc agtctgcccggctgacctca 240 aggcgactga agacaaagaa ggctgggctg gggccggtgg ccttgtggggccacgtggtg 300 gcctgctgtt ctgtccgaac agggctgtcg ttgtgaagcc attgcaccgagatgtccggg 360 ggaaagaagt tctggatcag gcacgtgaga atgactttgt ccttggtcttcggctccccc 420 tcgggcggca ggaacacgta cacctccggg gggaaacgcc ggccgggggctttggcaatg 480 gagcgcacga tgtccttggg caggtctgga tgggtcacct tgcattggtaggtctccccc 540 tcaacccagt cagtggcatc cacaggcagg gtggaagtga cggtgattgtcccgttgtac 600 tgagtcttct tgaccatcgg gtctgggtgc acagactccc cattctcccgcgaccaggtc 660 aggatcatgc cgtctgtgtt ggccaggtcc accaccaggc aggtgatcttgggcgacttg 720 tggacgtaca ggtcaagagg gctgggcggg ctcaagtagg tgctcacacctcgggggtca 780 gactctgtgc acttgcgagc gtggtcctca aaggtgaagc cttgataggtgacctggcaa 840 gtgtaggtct tttgggacac ccactcaccc tgcgtgatgt tgagctcgctgtgggtggag 900 gtcaccttgc cctcctgctt gccgggggca gtgtatggga atatgttcgtggccttctgc 960 ccatccacca gccaggtgac ctccatgtca cctgggacgt agccggagatgaggcacagg 1020 agctggatgg tgctaccggt gtcaccgagg gggttacagg aggagtggaagagcttcacg 1080 gtggggggaa tgaagttcat ggtacacgca ctgacggtct tgttgatggtgggggactcc 1140 gcgtgagcca cactgcaggt gaacttctgt ttggcccact cgcccgagacggtcacgtgg 1200 ctggtggtgg tgtagaggcc agaggtctcc tggagggtgg cggggagggtcacgacgctc 1260 ttgttcaggg accttgcatc ccaggtcaca gtcaccggca tcgggaagtagcccgtgacc 1320 aggcagccca gtgtcacgga cggggcagtg gcgatggtgc ctttgcagcaggtggccaag 1380 gggaagacga ggggggcctg gatggaggct gaggacac 1418 16 195DNA Felis catus 16 gcatatatta gtagtggagg taacacagac tacgcagactccgtgaaggg ccgattctcc 60 atctccagag acaacgccaa gaacacgctg tatctgcagatgaccagcct caagaccgag 120 gacacggcca catattactg tgcaagaggg actggtgtaataccggacta ctggggccag 180 ggagccctgg tgacg 195 17 64 PRT Felis catus 17Ala Tyr Ile Ser Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys 1 5 1015 Gly Arg Phe Ser Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr Leu 20 2530 Gln Met Thr Ser Leu Lys Thr Glu Asp Thr Ala Thr Tyr Tyr Cys Ala 35 4045 Arg Gly Thr Gly Val Ile Pro Asp Tyr Trp Gly Gln Gly Ala Leu Val 50 5560 18 195 DNA Felis catus 18 cgtcaccagg gctccctggc cccagtagtc cggtattacaccagtccctc ttgcacagta 60 atatgtggcc gtgtcctcgg tcttgaggct ggtcatctgcagatacagcg tgttcttggc 120 gttgtctctg gagatggaga atcggccctt cacggagtctgcgtagtctg tgttacctcc 180 actactaata tatgc 195 19 954 DNA Felis catusCDS (7)..(732) 19 ctcaaa atg agg ttc cct gct cag ctc ctg gga ctc atc atgctc tgg 48 Met Arg Phe Pro Ala Gln Leu Leu Gly Leu Ile Met Leu Trp 1 510 atc cca gga tcc agt ggg gat att gtg atg acg cag acc cct ctg tcc 96Ile Pro Gly Ser Ser Gly Asp Ile Val Met Thr Gln Thr Pro Leu Ser 15 20 2530 ctg tcc gtc acc cct gga gag cca gcc tca atc tcc tgc agg gcc agt 144Leu Ser Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ala Ser 35 40 45cag agc ctc ctg tac agt gat gga aat act tat ctg aat tgg tac ctg 192 GlnSer Leu Leu Tyr Ser Asp Gly Asn Thr Tyr Leu Asn Trp Tyr Leu 50 55 60 cagaag cca ggc cag tct cca cgg cgc ttg atc tat ctt gtt tcc aac 240 Gln LysPro Gly Gln Ser Pro Arg Arg Leu Ile Tyr Leu Val Ser Asn 65 70 75 cgg gactct ggg gtc cca gac agg ttc agt ggc agt ggg tca ggg aca 288 Arg Asp SerGly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr 80 85 90 gat ttc accctg aga atc agc agg gtg gag gct gac gac gtc ggt gtt 336 Asp Phe Thr LeuArg Ile Ser Arg Val Glu Ala Asp Asp Val Gly Val 95 100 105 110 tat tactgc ggt caa ggt tta cag cat cct ctc act ttc ggc cca ggt 384 Tyr Tyr CysGly Gln Gly Leu Gln His Pro Leu Thr Phe Gly Pro Gly 115 120 125 acc aagctg gag atc aaa cgg agt gat gct cag cca tct gtc ttt ctc 432 Thr Lys LeuGlu Ile Lys Arg Ser Asp Ala Gln Pro Ser Val Phe Leu 130 135 140 ttc caacca tct ctg gac gag tta cat aca gga agt gcc tct atc gtg 480 Phe Gln ProSer Leu Asp Glu Leu His Thr Gly Ser Ala Ser Ile Val 145 150 155 tgc atattg aat gac ttc tac ccc aaa gag gtc aat gtc aag tgg aaa 528 Cys Ile LeuAsn Asp Phe Tyr Pro Lys Glu Val Asn Val Lys Trp Lys 160 165 170 gtg gatggc gta gtc caa aac aaa ggc atc cag gag agc acc aca gag 576 Val Asp GlyVal Val Gln Asn Lys Gly Ile Gln Glu Ser Thr Thr Glu 175 180 185 190 cagaac agc aag gac agc acc tac agc ctc agc agc acc ctg acg atg 624 Gln AsnSer Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Met 195 200 205 tccagt acg gag tac caa agt cat gaa aag ttc tcc tgc gag gtc act 672 Ser SerThr Glu Tyr Gln Ser His Glu Lys Phe Ser Cys Glu Val Thr 210 215 220 cacaag agc ctg gcc tcc acc ctc gtc aag agc ttc aac agg agc gag 720 His LysSer Leu Ala Ser Thr Leu Val Lys Ser Phe Asn Arg Ser Glu 225 230 235 tgtcag aga gag tagcctagca ggcctcatca cctgtgcctc agtcccagac 772 Cys Gln ArgGlu 240 tctctgtctc cctcctcagg cctccggacc tttccccatc ggagacccacacctattgca 832 ggcccttgtc cccaccttac tgcctccccc tctttggctt taatcatgctaataatatat 892 gggggggaaa tgaataaata aagtgaatct ttgcaccagt gaaaaaaaaaaaaaaaaaaa 952 aa 954 20 242 PRT Felis catus 20 Met Arg Phe Pro Ala GlnLeu Leu Gly Leu Ile Met Leu Trp Ile Pro 1 5 10 15 Gly Ser Ser Gly AspIle Val Met Thr Gln Thr Pro Leu Ser Leu Ser 20 25 30 Val Thr Pro Gly GluPro Ala Ser Ile Ser Cys Arg Ala Ser Gln Ser 35 40 45 Leu Leu Tyr Ser AspGly Asn Thr Tyr Leu Asn Trp Tyr Leu Gln Lys 50 55 60 Pro Gly Gln Ser ProArg Arg Leu Ile Tyr Leu Val Ser Asn Arg Asp 65 70 75 80 Ser Gly Val ProAsp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe 85 90 95 Thr Leu Arg IleSer Arg Val Glu Ala Asp Asp Val Gly Val Tyr Tyr 100 105 110 Cys Gly GlnGly Leu Gln His Pro Leu Thr Phe Gly Pro Gly Thr Lys 115 120 125 Leu GluIle Lys Arg Ser Asp Ala Gln Pro Ser Val Phe Leu Phe Gln 130 135 140 ProSer Leu Asp Glu Leu His Thr Gly Ser Ala Ser Ile Val Cys Ile 145 150 155160 Leu Asn Asp Phe Tyr Pro Lys Glu Val Asn Val Lys Trp Lys Val Asp 165170 175 Gly Val Val Gln Asn Lys Gly Ile Gln Glu Ser Thr Thr Glu Gln Asn180 185 190 Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Met SerSer 195 200 205 Thr Glu Tyr Gln Ser His Glu Lys Phe Ser Cys Glu Val ThrHis Lys 210 215 220 Ser Leu Ala Ser Thr Leu Val Lys Ser Phe Asn Arg SerGlu Cys Gln 225 230 235 240 Arg Glu 21 954 DNA Felis catus 21 tttttttttttttttttttt tcactggtgc aaagattcac tttatttatt catttccccc 60 ccatatattattagcatgat taaagccaaa gagggggagg cagtaaggtg gggacaaggg 120 cctgcaataggtgtgggtct ccgatgggga aaggtccgga ggcctgagga gggagacaga 180 gagtctgggactgaggcaca ggtgatgagg cctgctaggc tactctctct gacactcgct 240 cctgttgaagctcttgacga gggtggaggc caggctcttg tgagtgacct cgcaggagaa 300 cttttcatgactttggtact ccgtactgga catcgtcagg gtgctgctga ggctgtaggt 360 gctgtccttgctgttctgct ctgtggtgct ctcctggatg cctttgtttt ggactacgcc 420 atccactttccacttgacat tgacctcttt ggggtagaag tcattcaata tgcacacgat 480 agaggcacttcctgtatgta actcgtccag agatggttgg aagagaaaga cagatggctg 540 agcatcactccgtttgatct ccagcttggt acctgggccg aaagtgagag gatgctgtaa 600 accttgaccgcagtaataaa caccgacgtc gtcagcctcc accctgctga ttctcagggt 660 gaaatctgtccctgacccac tgccactgaa cctgtctggg accccagagt cccggttgga 720 aacaagatagatcaagcgcc gtggagactg gcctggcttc tgcaggtacc aattcagata 780 agtatttccatcactgtaca ggaggctctg actggccctg caggagattg aggctggctc 840 tccaggggtgacggacaggg acagaggggt ctgcgtcatc acaatatccc cactggatcc 900 tgggatccagagcatgatga gtcccaggag ctgagcaggg aacctcattt tgag 954 22 6 DNA Feliscatus 22 aataaa 6 23 726 DNA Felis catus 23 atgaggttcc ctgctcagctcctgggactc atcatgctct ggatcccagg atccagtggg 60 gatattgtga tgacgcagacccctctgtcc ctgtccgtca cccctggaga gccagcctca 120 atctcctgca gggccagtcagagcctcctg tacagtgatg gaaatactta tctgaattgg 180 tacctgcaga agccaggccagtctccacgg cgcttgatct atcttgtttc caaccgggac 240 tctggggtcc cagacaggttcagtggcagt gggtcaggga cagatttcac cctgagaatc 300 agcagggtgg aggctgacgacgtcggtgtt tattactgcg gtcaaggttt acagcatcct 360 ctcactttcg gcccaggtaccaagctggag atcaaacgga gtgatgctca gccatctgtc 420 tttctcttcc aaccatctctggacgagtta catacaggaa gtgcctctat cgtgtgcata 480 ttgaatgact tctaccccaaagaggtcaat gtcaagtgga aagtggatgg cgtagtccaa 540 aacaaaggca tccaggagagcaccacagag cagaacagca aggacagcac ctacagcctc 600 agcagcaccc tgacgatgtccagtacggag taccaaagtc atgaaaagtt ctcctgcgag 660 gtcactcaca agagcctggcctccaccctc gtcaagagct tcaacaggag cgagtgtcag 720 agagag 726 24 726 DNAFelis catus 24 ctctctctga cactcgctcc tgttgaagct cttgacgagg gtggaggccaggctcttgtg 60 agtgacctcg caggagaact tttcatgact ttggtactcc gtactggacatcgtcagggt 120 gctgctgagg ctgtaggtgc tgtccttgct gttctgctct gtggtgctctcctggatgcc 180 tttgttttgg actacgccat ccactttcca cttgacattg acctctttggggtagaagtc 240 attcaatatg cacacgatag aggcacttcc tgtatgtaac tcgtccagagatggttggaa 300 gagaaagaca gatggctgag catcactccg tttgatctcc agcttggtacctgggccgaa 360 agtgagagga tgctgtaaac cttgaccgca gtaataaaca ccgacgtcgtcagcctccac 420 cctgctgatt ctcagggtga aatctgtccc tgacccactg ccactgaacctgtctgggac 480 cccagagtcc cggttggaaa caagatagat caagcgccgt ggagactggcctggcttctg 540 caggtaccaa ttcagataag tatttccatc actgtacagg aggctctgactggccctgca 600 ggagattgag gctggctctc caggggtgac ggacagggac agaggggtctgcgtcatcac 660 aatatcccca ctggatcctg ggatccagag catgatgagt cccaggagctgagcagggaa 720 cctcat 726 25 666 DNA Felis catus 25 gatattgtgatgacgcagac ccctctgtcc ctgtccgtca cccctggaga gccagcctca 60 atctcctgcagggccagtca gagcctcctg tacagtgatg gaaatactta tctgaattgg 120 tacctgcagaagccaggcca gtctccacgg cgcttgatct atcttgtttc caaccgggac 180 tctggggtcccagacaggtt cagtggcagt gggtcaggga cagatttcac cctgagaatc 240 agcagggtggaggctgacga cgtcggtgtt tattactgcg gtcaaggttt acagcatcct 300 ctcactttcggcccaggtac caagctggag atcaaacgga gtgatgctca gccatctgtc 360 tttctcttccaaccatctct ggacgagtta catacaggaa gtgcctctat cgtgtgcata 420 ttgaatgacttctaccccaa agaggtcaat gtcaagtgga aagtggatgg cgtagtccaa 480 aacaaaggcatccaggagag caccacagag cagaacagca aggacagcac ctacagcctc 540 agcagcaccctgacgatgtc cagtacggag taccaaagtc atgaaaagtt ctcctgcgag 600 gtcactcacaagagcctggc ctccaccctc gtcaagagct tcaacaggag cgagtgtcag 660 agagag 666 26222 PRT Felis catus 26 Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu SerVal Thr Pro Gly 1 5 10 15 Glu Pro Ala Ser Ile Ser Cys Arg Ala Ser GlnSer Leu Leu Tyr Ser 20 25 30 Asp Gly Asn Thr Tyr Leu Asn Trp Tyr Leu GlnLys Pro Gly Gln Ser 35 40 45 Pro Arg Arg Leu Ile Tyr Leu Val Ser Asn ArgAsp Ser Gly Val Pro 50 55 60 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr AspPhe Thr Leu Arg Ile 65 70 75 80 Ser Arg Val Glu Ala Asp Asp Val Gly ValTyr Tyr Cys Gly Gln Gly 85 90 95 Leu Gln His Pro Leu Thr Phe Gly Pro GlyThr Lys Leu Glu Ile Lys 100 105 110 Arg Ser Asp Ala Gln Pro Ser Val PheLeu Phe Gln Pro Ser Leu Asp 115 120 125 Glu Leu His Thr Gly Ser Ala SerIle Val Cys Ile Leu Asn Asp Phe 130 135 140 Tyr Pro Lys Glu Val Asn ValLys Trp Lys Val Asp Gly Val Val Gln 145 150 155 160 Asn Lys Gly Ile GlnGlu Ser Thr Thr Glu Gln Asn Ser Lys Asp Ser 165 170 175 Thr Tyr Ser LeuSer Ser Thr Leu Thr Met Ser Ser Thr Glu Tyr Gln 180 185 190 Ser His GluLys Phe Ser Cys Glu Val Thr His Lys Ser Leu Ala Ser 195 200 205 Thr LeuVal Lys Ser Phe Asn Arg Ser Glu Cys Gln Arg Glu 210 215 220 27 666 DNAFelis catus 27 ctctctctga cactcgctcc tgttgaagct cttgacgagg gtggaggccaggctcttgtg 60 agtgacctcg caggagaact tttcatgact ttggtactcc gtactggacatcgtcagggt 120 gctgctgagg ctgtaggtgc tgtccttgct gttctgctct gtggtgctctcctggatgcc 180 tttgttttgg actacgccat ccactttcca cttgacattg acctctttggggtagaagtc 240 attcaatatg cacacgatag aggcacttcc tgtatgtaac tcgtccagagatggttggaa 300 gagaaagaca gatggctgag catcactccg tttgatctcc agcttggtacctgggccgaa 360 agtgagagga tgctgtaaac cttgaccgca gtaataaaca ccgacgtcgtcagcctccac 420 cctgctgatt ctcagggtga aatctgtccc tgacccactg ccactgaacctgtctgggac 480 cccagagtcc cggttggaaa caagatagat caagcgccgt ggagactggcctggcttctg 540 caggtaccaa ttcagataag tatttccatc actgtacagg aggctctgactggccctgca 600 ggagattgag gctggctctc caggggtgac ggacagggac agaggggtctgcgtcatcac 660 aatatc 666 28 1613 DNA Felis catus 28 gcatatattagtagtggagg taacacagac tacgcagact ccgtgaaggg ccgattctcc 60 atctccagagacaacgccaa gaacacgctg tatctgcaga tgaccagcct caagaccgag 120 gacacggccacatattactg tgcaagaggg actggtgtaa taccggacta ctggggccag 180 ggagccctggtgacggtgtc ctcaacctcc atccaggccc ccctcgtctt ccccttggcc 240 acctgctgcaaaggcaccat cgccactgcc ccgtccgtga cactgggctg cctggtcacg 300 ggctacttcccgatgccggt gactgtgacc tgggatgcaa ggtccctgaa caagagcgtc 360 gtgaccctccccgccaccct ccaggagacc tctggcctct acaccaccac cagccacgtg 420 accgtctcgggcgagtgggc caaacagaag ttcacctgca gtgtggctca cgcggagtcc 480 cccaccatcaacaagaccgt cagtgcgtgt accatgaact tcattccccc caccgtgaag 540 ctcttccactcctcctgtaa ccccctcggt gacaccggta gcaccatcca gctcctgtgc 600 ctcatctccggctacgtccc aggtgacatg gaggtcacct ggctggtgga tgggcagaag 660 gccacgaacatattcccata cactgccccc ggcaagcagg agggcaaggt gacctccacc 720 cacagcgagctcaacatcac gcagggtgag tgggtgtccc aaaagaccta cacttgccag 780 gtcacctatcaaggcttcac ctttgaggac cacgctcgca agtgcacaga gtctgacccc 840 cgaggtgtgagcacctactt gagcccgccc agccctcttg acctgtacgt ccacaagtcg 900 cccaagatcacctgcctggt ggtggacctg gccaacacag acggcatgat cctgacctgg 960 tcgcgggagaatggggagtc tgtgcaccca gacccgatgg tcaagaagac tcagtacaac 1020 gggacaatcaccgtcacttc caccctgcct gtggatgcca ctgactgggt tgagggggag 1080 acctaccaatgcaaggtgac ccatccagac ctgcccaagg acatcgtgcg ctccattgcc 1140 aaagcccccggccggcgttt ccccccggag gtgtacgtgt tcctgccgcc cgagggggag 1200 ccgaagaccaaggacaaagt cattctcacg tgcctgatcc agaacttctt tcccccggac 1260 atctcggtgcaatggcttca caacgacagc cctgttcgga cagaacagca ggccaccacg 1320 tggccccacaaggccaccgg ccccagccca gccttctttg tcttcagtcg ccttgaggtc 1380 agccgggcagactgggagca gagggatgtg ttcacctgcc aagtggtgca cgaggcgctg 1440 cctggctttaggacgctcaa gaaatccgtg tccaaaaacc ctggtaaatg atgcccaccc 1500 ctccccccagagctccatcc tgctggggcg ggggaggggc cggccggacc tgccggtctg 1560 ttgttgtcaataaacactgc agtgcctgcc tcagaaaaaa aaaaaaaaaa aaa 1613 29 496 PRT Feliscatus 29 Ala Tyr Ile Ser Ser Gly Gly Asn Thr Asp Tyr Ala Asp Ser Val Lys1 5 10 15 Gly Arg Phe Ser Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu TyrLeu 20 25 30 Gln Met Thr Ser Leu Lys Thr Glu Asp Thr Ala Thr Tyr Tyr CysAla 35 40 45 Arg Gly Thr Gly Val Ile Pro Asp Tyr Trp Gly Gln Gly Ala LeuVal 50 55 60 Thr Val Ser Ser Thr Ser Ile Gln Ala Pro Leu Val Phe Pro LeuAla 65 70 75 80 Thr Cys Cys Lys Gly Thr Ile Ala Thr Ala Pro Ser Val ThrLeu Gly 85 90 95 Cys Leu Val Thr Gly Tyr Phe Pro Met Pro Val Thr Val ThrTrp Asp 100 105 110 Ala Arg Ser Leu Asn Lys Ser Val Val Thr Leu Pro AlaThr Leu Gln 115 120 125 Glu Thr Ser Gly Leu Tyr Thr Thr Thr Ser His ValThr Val Ser Gly 130 135 140 Glu Trp Ala Lys Gln Lys Phe Thr Cys Ser ValAla His Ala Glu Ser 145 150 155 160 Pro Thr Ile Asn Lys Thr Val Ser AlaCys Thr Met Asn Phe Ile Pro 165 170 175 Pro Thr Val Lys Leu Phe His SerSer Cys Asn Pro Leu Gly Asp Thr 180 185 190 Gly Ser Thr Ile Gln Leu LeuCys Leu Ile Ser Gly Tyr Val Pro Gly 195 200 205 Asp Met Glu Val Thr TrpLeu Val Asp Gly Gln Lys Ala Thr Asn Ile 210 215 220 Phe Pro Tyr Thr AlaPro Gly Lys Gln Glu Gly Lys Val Thr Ser Thr 225 230 235 240 His Ser GluLeu Asn Ile Thr Gln Gly Glu Trp Val Ser Gln Lys Thr 245 250 255 Tyr ThrCys Gln Val Thr Tyr Gln Gly Phe Thr Phe Glu Asp His Ala 260 265 270 ArgLys Cys Thr Glu Ser Asp Pro Arg Gly Val Ser Thr Tyr Leu Ser 275 280 285Pro Pro Ser Pro Leu Asp Leu Tyr Val His Lys Ser Pro Lys Ile Thr 290 295300 Cys Leu Val Val Asp Leu Ala Asn Thr Asp Gly Met Ile Leu Thr Trp 305310 315 320 Ser Arg Glu Asn Gly Glu Ser Val His Pro Asp Pro Met Val LysLys 325 330 335 Thr Gln Tyr Asn Gly Thr Ile Thr Val Thr Ser Thr Leu ProVal Asp 340 345 350 Ala Thr Asp Trp Val Glu Gly Glu Thr Tyr Gln Cys LysVal Thr His 355 360 365 Pro Asp Leu Pro Lys Asp Ile Val Arg Ser Ile AlaLys Ala Pro Gly 370 375 380 Arg Arg Phe Pro Pro Glu Val Tyr Val Phe LeuPro Pro Glu Gly Glu 385 390 395 400 Pro Lys Thr Lys Asp Lys Val Ile LeuThr Cys Leu Ile Gln Asn Phe 405 410 415 Phe Pro Pro Asp Ile Ser Val GlnTrp Leu His Asn Asp Ser Pro Val 420 425 430 Arg Thr Glu Gln Gln Ala ThrThr Trp Pro His Lys Ala Thr Gly Pro 435 440 445 Ser Pro Ala Phe Phe ValPhe Ser Arg Leu Glu Val Ser Arg Ala Asp 450 455 460 Trp Glu Gln Arg AspVal Phe Thr Cys Gln Val Val His Glu Ala Leu 465 470 475 480 Pro Gly PheArg Thr Leu Lys Lys Ser Val Ser Lys Asn Pro Gly Lys 485 490 495 30 1613DNA Felis catus 30 tttttttttt tttttttttc tgaggcaggc actgcagtgtttattgacaa caacagaccg 60 gcaggtccgg ccggcccctc ccccgcccca gcaggatggagctctggggg gaggggtggg 120 catcatttac cagggttttt ggacacggat ttcttgagcgtcctaaagcc aggcagcgcc 180 tcgtgcacca cttggcaggt gaacacatcc ctctgctcccagtctgcccg gctgacctca 240 aggcgactga agacaaagaa ggctgggctg gggccggtggccttgtgggg ccacgtggtg 300 gcctgctgtt ctgtccgaac agggctgtcg ttgtgaagccattgcaccga gatgtccggg 360 ggaaagaagt tctggatcag gcacgtgaga atgactttgtccttggtctt cggctccccc 420 tcgggcggca ggaacacgta cacctccggg gggaaacgccggccgggggc tttggcaatg 480 gagcgcacga tgtccttggg caggtctgga tgggtcaccttgcattggta ggtctccccc 540 tcaacccagt cagtggcatc cacaggcagg gtggaagtgacggtgattgt cccgttgtac 600 tgagtcttct tgaccatcgg gtctgggtgc acagactccccattctcccg cgaccaggtc 660 aggatcatgc cgtctgtgtt ggccaggtcc accaccaggcaggtgatctt gggcgacttg 720 tggacgtaca ggtcaagagg gctgggcggg ctcaagtaggtgctcacacc tcgggggtca 780 gactctgtgc acttgcgagc gtggtcctca aaggtgaagccttgataggt gacctggcaa 840 gtgtaggtct tttgggacac ccactcaccc tgcgtgatgttgagctcgct gtgggtggag 900 gtcaccttgc cctcctgctt gccgggggca gtgtatgggaatatgttcgt ggccttctgc 960 ccatccacca gccaggtgac ctccatgtca cctgggacgtagccggagat gaggcacagg 1020 agctggatgg tgctaccggt gtcaccgagg gggttacaggaggagtggaa gagcttcacg 1080 gtggggggaa tgaagttcat ggtacacgca ctgacggtcttgttgatggt gggggactcc 1140 gcgtgagcca cactgcaggt gaacttctgt ttggcccactcgcccgagac ggtcacgtgg 1200 ctggtggtgg tgtagaggcc agaggtctcc tggagggtggcggggagggt cacgacgctc 1260 ttgttcaggg accttgcatc ccaggtcaca gtcaccggcatcgggaagta gcccgtgacc 1320 aggcagccca gtgtcacgga cggggcagtg gcgatggtgcctttgcagca ggtggccaag 1380 gggaagacga ggggggcctg gatggaggtt gaggacaccgtcaccagggc tccctggccc 1440 cagtagtccg gtattacacc agtccctctt gcacagtaatatgtggccgt gtcctcggtc 1500 ttgaggctgg tcatctgcag atacagcgtg ttcttggcgttgtctctgga gatggagaat 1560 cggcccttca cggagtctgc gtagtctgtg ttacctccactactaatata tgc 1613 31 1488 DNA Felis catus 31 gcatatatta gtagtggaggtaacacagac tacgcagact ccgtgaaggg ccgattctcc 60 atctccagag acaacgccaagaacacgctg tatctgcaga tgaccagcct caagaccgag 120 gacacggcca catattactgtgcaagaggg actggtgtaa taccggacta ctggggccag 180 ggagccctgg tgacggtgtcctcaacctcc atccaggccc ccctcgtctt ccccttggcc 240 acctgctgca aaggcaccatcgccactgcc ccgtccgtga cactgggctg cctggtcacg 300 ggctacttcc cgatgccggtgactgtgacc tgggatgcaa ggtccctgaa caagagcgtc 360 gtgaccctcc ccgccaccctccaggagacc tctggcctct acaccaccac cagccacgtg 420 accgtctcgg gcgagtgggccaaacagaag ttcacctgca gtgtggctca cgcggagtcc 480 cccaccatca acaagaccgtcagtgcgtgt accatgaact tcattccccc caccgtgaag 540 ctcttccact cctcctgtaaccccctcggt gacaccggta gcaccatcca gctcctgtgc 600 ctcatctccg gctacgtcccaggtgacatg gaggtcacct ggctggtgga tgggcagaag 660 gccacgaaca tattcccatacactgccccc ggcaagcagg agggcaaggt gacctccacc 720 cacagcgagc tcaacatcacgcagggtgag tgggtgtccc aaaagaccta cacttgccag 780 gtcacctatc aaggcttcacctttgaggac cacgctcgca agtgcacaga gtctgacccc 840 cgaggtgtga gcacctacttgagcccgccc agccctcttg acctgtacgt ccacaagtcg 900 cccaagatca cctgcctggtggtggacctg gccaacacag acggcatgat cctgacctgg 960 tcgcgggaga atggggagtctgtgcaccca gacccgatgg tcaagaagac tcagtacaac 1020 gggacaatca ccgtcacttccaccctgcct gtggatgcca ctgactgggt tgagggggag 1080 acctaccaat gcaaggtgacccatccagac ctgcccaagg acatcgtgcg ctccattgcc 1140 aaagcccccg gccggcgtttccccccggag gtgtacgtgt tcctgccgcc cgagggggag 1200 ccgaagacca aggacaaagtcattctcacg tgcctgatcc agaacttctt tcccccggac 1260 atctcggtgc aatggcttcacaacgacagc cctgttcgga cagaacagca ggccaccacg 1320 tggccccaca aggccaccggccccagccca gccttctttg tcttcagtcg ccttgaggtc 1380 agccgggcag actgggagcagagggatgtg ttcacctgcc aagtggtgca cgaggcgctg 1440 cctggcttta ggacgctcaagaaatccgtg tccaaaaacc ctggtaaa 1488 32 1488 DNA Felis catus 32tttaccaggg tttttggaca cggatttctt gagcgtccta aagccaggca gcgcctcgtg 60caccacttgg caggtgaaca catccctctg ctcccagtct gcccggctga cctcaaggcg 120actgaagaca aagaaggctg ggctggggcc ggtggccttg tggggccacg tggtggcctg 180ctgttctgtc cgaacagggc tgtcgttgtg aagccattgc accgagatgt ccgggggaaa 240gaagttctgg atcaggcacg tgagaatgac tttgtccttg gtcttcggct ccccctcggg 300cggcaggaac acgtacacct ccggggggaa acgccggccg ggggctttgg caatggagcg 360cacgatgtcc ttgggcaggt ctggatgggt caccttgcat tggtaggtct ccccctcaac 420ccagtcagtg gcatccacag gcagggtgga agtgacggtg attgtcccgt tgtactgagt 480cttcttgacc atcgggtctg ggtgcacaga ctccccattc tcccgcgacc aggtcaggat 540catgccgtct gtgttggcca ggtccaccac caggcaggtg atcttgggcg acttgtggac 600gtacaggtca agagggctgg gcgggctcaa gtaggtgctc acacctcggg ggtcagactc 660tgtgcacttg cgagcgtggt cctcaaaggt gaagccttga taggtgacct ggcaagtgta 720ggtcttttgg gacacccact caccctgcgt gatgttgagc tcgctgtggg tggaggtcac 780cttgccctcc tgcttgccgg gggcagtgta tgggaatatg ttcgtggcct tctgcccatc 840caccagccag gtgacctcca tgtcacctgg gacgtagccg gagatgaggc acaggagctg 900gatggtgcta ccggtgtcac cgagggggtt acaggaggag tggaagagct tcacggtggg 960gggaatgaag ttcatggtac acgcactgac ggtcttgttg atggtggggg actccgcgtg 1020agccacactg caggtgaact tctgtttggc ccactcgccc gagacggtca cgtggctggt 1080ggtggtgtag aggccagagg tctcctggag ggtggcgggg agggtcacga cgctcttgtt 1140cagggacctt gcatcccagg tcacagtcac cggcatcggg aagtagcccg tgaccaggca 1200gcccagtgtc acggacgggg cagtggcgat ggtgcctttg cagcaggtgg ccaaggggaa 1260gacgaggggg gcctggatgg aggttgagga caccgtcacc agggctccct ggccccagta 1320gtccggtatt acaccagtcc ctcttgcaca gtaatatgtg gccgtgtcct cggtcttgag 1380gctggtcatc tgcagataca gcgtgttctt ggcgttgtct ctggagatgg agaatcggcc 1440cttcacggag tctgcgtagt ctgtgttacc tccactacta atatatgc 1488 33 1293 DNAFelis catus 33 gtgtcctcag cctccatcca ggcccccctc gtcttcccct tggccacctgctgcaaaggc 60 accatcgcca ctgccccgtc cgtgacactg ggctgcctgg tcacgggctacttcccgatg 120 ccggtgactg tgacctggga tgcaaggtcc ctgaacaaga gcgtcgtgaccctccccgcc 180 accctccagg agacctctgg cctctacacc accaccagcc acgtgaccgtctcgggcgag 240 tgggccaaac agaagttcac ctgcagtgtg gctcacgcgg agtcccccaccatcaacaag 300 accgtcagtg cgtgtaccat gaacttcatt ccccccaccg tgaagctcttccactcctcc 360 tgtaaccccc tcggtgacac cggtagcacc atccagctcc tgtgcctcatctccggctac 420 gtcccaggtg acatggaggt cacctggctg gtggatgggc agaaggccacgaacatattc 480 ccatacactg cccccggcaa gcaggagggc aaggtgacct ccacccacagcgagctcaac 540 atcacgcagg gtgagtgggt gtcccaaaag acctacactt gccaggtcacctatcaaggc 600 ttcacctttg aggaccacgc tcgcaagtgc acagagtctg acccccgaggtgtgagcacc 660 tacttgagcc cgcccagccc tcttgacctg tacgtccaca agtcgcccaagatcacctgc 720 ctggtggtgg acctggccaa cacagacggc atgatcctga cctggtcgcgggagaatggg 780 gagtctgtgc acccagaccc gatggtcaag aagactcagt acaacgggacaatcaccgtc 840 acttccaccc tgcctgtgga tgccactgac tgggttgagg gggagacctaccaatgcaag 900 gtgacccatc cagacctgcc caaggacatc gtgcgctcca ttgccaaagcccccggccgg 960 cgtttccccc cggaggtgta cgtgttcctg ccgcccgagg gggagccgaagaccaaggac 1020 aaagtcattc tcacgtgcct gatccagaac ttctttcccc cggacatctcggtgcaatgg 1080 cttcacaacg acagccctgt tcggacagaa cagcaggcca ccacgtggccccacaaggcc 1140 accggcccca gcccagcctt ctttgtcttc agtcgccttg aggtcagccgggcagactgg 1200 gagcagaggg atgtgttcac ctgccaagtg gtgcacgagg cgctgcctggctttaggacg 1260 ctcaagaaat ccgtgtccaa aaaccctggt aaa 1293 34 1293 DNAFelis catus 34 tttaccaggg tttttggaca cggatttctt gagcgtccta aagccaggcagcgcctcgtg 60 caccacttgg caggtgaaca catccctctg ctcccagtct gcccggctgacctcaaggcg 120 actgaagaca aagaaggctg ggctggggcc ggtggccttg tggggccacgtggtggcctg 180 ctgttctgtc cgaacagggc tgtcgttgtg aagccattgc accgagatgtccgggggaaa 240 gaagttctgg atcaggcacg tgagaatgac tttgtccttg gtcttcggctccccctcggg 300 cggcaggaac acgtacacct ccggggggaa acgccggccg ggggctttggcaatggagcg 360 cacgatgtcc ttgggcaggt ctggatgggt caccttgcat tggtaggtctccccctcaac 420 ccagtcagtg gcatccacag gcagggtgga agtgacggtg attgtcccgttgtactgagt 480 cttcttgacc atcgggtctg ggtgcacaga ctccccattc tcccgcgaccaggtcaggat 540 catgccgtct gtgttggcca ggtccaccac caggcaggtg atcttgggcgacttgtggac 600 gtacaggtca agagggctgg gcgggctcaa gtaggtgctc acacctcgggggtcagactc 660 tgtgcacttg cgagcgtggt cctcaaaggt gaagccttga taggtgacctggcaagtgta 720 ggtcttttgg gacacccact caccctgcgt gatgttgagc tcgctgtgggtggaggtcac 780 cttgccctcc tgcttgccgg gggcagtgta tgggaatatg ttcgtggccttctgcccatc 840 caccagccag gtgacctcca tgtcacctgg gacgtagccg gagatgaggcacaggagctg 900 gatggtgcta ccggtgtcac cgagggggtt acaggaggag tggaagagcttcacggtggg 960 gggaatgaag ttcatggtac acgcactgac ggtcttgttg atggtgggggactccgcgtg 1020 agccacactg caggtgaact tctgtttggc ccactcgccc gagacggtcacgtggctggt 1080 ggtggtgtag aggccagagg tctcctggag ggtggcgggg agggtcacgacgctcttgtt 1140 cagggacctt gcatcccagg tcacagtcac cggcatcggg aagtagcccgtgaccaggca 1200 gcccagtgtc acggacgggg cagtggcgat ggtgcctttg cagcaggtggccaaggggaa 1260 gacgaggggg gcctggatgg aggctgagga cac 1293

What is claimed is:
 1. An isolated nucleic acid molecule encoding aportion of a feline IgE heavy chain protein, wherein said nucleic acidmolecule comprises a nucleic acid sequence selected from the groupconsisting of: (a) a nucleic acid sequence which has more than 82%identity to a nucleic acid sequence selected from the group consistingof: SEQ ID NO 1; and SEQ ID NO 28, wherein said identity can bedetermined using the DNAsis computer program and default parameters; (b)a nucleic acid sequence which encodes a feline heavy chain protein whichhas more than 76% identity to an amino acid sequence selected from thegroup consisting of: SEQ ID NO 2; and SEQ ID NO 29, wherein saididentity can be determined using the DNAsis computer program and defaultparameters; (c) a nucleic acid sequence which encodes and a feline heavychain protein encoded by an allelic variant of a nucleic acid sequenceselected from the group consisting of: SEQ ID NO 1; and SEQ ID NO 28;and (d) a nucleic acid sequence which has more than 90% identity to anucleic acid sequence selected from the group consisting of: SEQ ID NO3; SEQ ID NO 4; SEQ ID NO 6; SEQ ID NO 7; SEQ ID NO 9; SEQ ID NO 10; SEQID NO 12; SEQ ID NO 13; SEQ ID NO 15; SEQ ID NO 16; SEQ ID NO 18; andSEQ ID NO 31; and (e) a nucleic acid molecule fully complementary to anucleic acid molecule selected from the group consisting of: a nucleicacid molecule of (a); a nucleic acid molecule of (b); and a nucleic acidmolecule of (c).
 2. An isolated nucleic acid molecule of claim 1,wherein said nucleic acid molecule encodes a feline IgE Fce receptorbinding region and which comprises SEQ ID NO
 4. 3. An isolated nucleicacid molecule of claim 1, wherein said nucleic acid molecule encodes afeline IgE Fce receptor binding region and which comprises SEQ ID NO 7.4. An isolated nucleic acid molecule of claim 1, wherein said nucleicacid molecule encodes a feline IgE Fce receptor binding region and whichcomprises SEQ ID NO
 10. 5. An isolated nucleic acid molecule of claim 1,wherein said nucleic acid molecule encodes a feline IgE constant regionand comprises SEQ ID NO
 13. 6. An expression vector comprising a nucleicacid molecule of claim
 1. 7. A recombinant cell transformed with avector comprising a nucleic acid molecule of claim
 1. 8. An isolatedprotein encoding a portion of a feline IgE heavy chain molecule, whereinsaid protein comprises an amino acid sequence selected from the groupconsisting of: (a) an amino acid sequence encoded by a nucleic acidsequence which has more than 82% identity to a nucleic acid sequenceselected from the group consisting of: SEQ ID NO 1; and SEQ ID NO: 28,wherein said identity can be determined using the DNAsis computerprogram and default parameters; (b) an amino acid sequence which hasmore than 76% identity to an amino acid sequence selected from the groupconsisting of: SEQ ID NO 2; and SEQ ID NO: 29, wherein said identity canbe determined using the DNAsis computer program and default parameters;(c) an amino acid sequence encoded by an allelic variant of a nucleicacid sequence selected from the group consisting of: SEQ ID NO 1; andSEQ ID NO: 28; and (d) an amino acid sequence encoded by a a nucleicacid sequence which has more than 90% identity to a nucleic acidsequence selected from the group consisting of: SEQ ID NO 3; SEQ ID NO4; SEQ ID NO 6; SEQ ID NO 7; SEQ ID NO 9; SEQ ID NO 10; SEQ ID NO 12;SEQ ID NO 13; SEQ ID NO 15; SEQ ID NO 16; SEQ ID NO 18; and SEQ ID NO31.
 9. An isolated protein of claim 8, wherein said protein is capableof binding to Fce receptor and comprises SEQ ID NO
 11. 10. An isolatedprotein of claim 8, wherein said protein is capable of binding to Fcereceptor and comprises SEQ ID NO
 8. 11. An isolated protein of claim 8,wherein said protein is capable of binding to Fce receptor and comprisesSEQ ID NO
 5. 12. An isolated protein of claim 8, wherein said protein iscapable of binding to Fce receptor and comprises SEQ ID NO
 14. 13. Anantibody selective for a protein of claim
 8. 14. An antibody of claim13, which i s selected from the group consisting of: H-100; H-101;H-102; H-103; and H-106.
 15. A therapeutic composition useful forinhibiting an immune response to feline IgE, wherein said therapeuticcomposition is selected from the group consisting of: (a) a nucleic acidmolecule of claim 1; (b) a protein encoded by a nucleic acid of (a); (c)an inhibitor of a nucleic acid of (a); and (d) an inhibitor of a proteinof (b).
 16. A therapeutic composition useful for inhibiting an immuneresponse to feline IgE, comprising an antibody of claim
 13. 17. A methodto identify the ability of a test compound to interfere with IgE/Fcereceptor interaction, comprising: contacting the test compound with aprotein of claim 8; and determining whether the test compound and saidprotein interact.
 18. A method for inhibiting an immune response tofeline IgE, comprising administering a therapeutic composition of claim15.
 19. A diagnostic kit, comprising a container comprising at least onecomposition selected from the group consisting of: (a) a nucleic acidmolecule of claim 1; (b) a protein encoded by a nucleic acid of (a); (c)an inhibitor of a nucleic acid of (a); and (d) an inhibitor of a proteinof (b).
 20. An isolated nucleic acid molecule encoding a portion of afeline IgE light chain protein, wherein said nucleic acid moleculecomprises a nucleic acid sequence selected from the group consisting of:(a) a nucleic acid sequence which has more than 84% identity to SEQ IDNO 19, and wherein said identity can be determined using the DNAsiscomputer program and default parameters; (b) a nucleic acid sequencewhich encodes a feline heavy chain protein selected from the groupconsisting of: a feline heavy chain protein which has more than 61%identity to SEQ ID NO 20, wherein said identity can be determined usingthe DNAsis computer program and default parameters; and a feline heavychain protein encoded by an allelic variant of SEQ ID NO 19; (c) anucleic acid sequence which has more than 95% identity to a nucleic acidsequence selected from the group consisting of: SEQ ID NO 23; and SEQ IDNO 25; and (d) a nucleic acid molecule fully complementary to a nucleicacid molecule selected from the group consisting of: a nucleic acidmolecule of (a); a nucleic acid molecule of (b); and a nucleic acidmolecule of (c).
 21. An isolated protein encoding a portion of a felineIgE light chain molecule, wherein said protein comprises an amino acidsequence selected from the group consisting of: (a) an amino acidsequence encoded by a nucleic acid sequence which has more than 84%identity to SEQ ID NO 19, and wherein said identity can be determinedusing the DNAsis computer program and default parameters; (b) an aminoacid sequence which has more than 61% identity to SEQ ID NO 20, whereinsaid identity can be determined using the DNAsis computer program anddefault parameters; (c) an amino acid sequence encoded by an allelicvariant of SEQ ID NO 19; and (d) a nucleic acid sequence which has morethan 95% identity to a nucleic acid sequence selected from the groupconsisting of: SEQ ID NO 23; and SEQ ID NO 25.